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|
||
<span class="search-input-link user-guide-link" role="navigation" aria-label="User guide">
|
||
<a href="/help/"
|
||
data-ga-category="featured_link"
|
||
data-ga-action="user_guide">
|
||
User Guide
|
||
</a>
|
||
</span>
|
||
|
||
</div>
|
||
</div>
|
||
|
||
|
||
</form>
|
||
|
||
|
||
<div id="article-page-header">
|
||
<div id="article-top-actions-bar" class="article-top-actions-bar">
|
||
<div class="inner-wrap">
|
||
<div class="actions-container">
|
||
<div class="return-to-search">
|
||
|
||
|
||
</div>
|
||
|
||
<div class="multiple-results-actions " role="region" aria-label="save, email, send to">
|
||
<button id="save-results-panel-trigger"
|
||
type="button"
|
||
class="save-results save-results-panel-trigger"
|
||
aria-expanded="false"
|
||
aria-controls="save-action-panel"
|
||
data-ga-category="save_share"
|
||
data-ga-action="save"
|
||
data-ga-label="open">
|
||
Save
|
||
</button><button id="email-results-panel-trigger"
|
||
type="button"
|
||
class="email-results email-results-panel-trigger"
|
||
aria-expanded="false"
|
||
aria-controls="email-action-panel"
|
||
data-ga-category="save_share"
|
||
data-ga-action="email"
|
||
data-ga-label="open">
|
||
Email
|
||
</button><div class="more-actions dropdown-block"><button id="more-actions-trigger"
|
||
type="button"
|
||
class="trigger more-actions-trigger"
|
||
ref="linksrc=show_moreactions_btn"
|
||
aria-label="Send to"
|
||
aria-controls="more-actions-dropdown">Send to</button><div id="more-actions-dropdown" class="dropdown dropdown-container" hidden><div class="content"><ul class="more-actions-links"><li><a class="clipboard-panel-trigger dropdown-block-link "
|
||
href="#"
|
||
role="button"
|
||
data-ga-category="save_share"
|
||
data-ga-action="clipboard"
|
||
data-ga-label="send">
|
||
Clipboard
|
||
</a></li><li><a role="button" class="dropdown-block-link bibliography-trigger-target" href="https://account.ncbi.nlm.nih.gov/?back_url=https%3A%2F%2Fpubmed.ncbi.nlm.nih.gov%2F28031477%2F%23open-bibliography-panel">My Bibliography</a></li><li><a role="button" class="dropdown-block-link collections-trigger-target" href="https://account.ncbi.nlm.nih.gov/?back_url=https%3A%2F%2Fpubmed.ncbi.nlm.nih.gov%2F28031477%2F%23open-collections-panel">Collections</a></li><li><a role="button" class="citation-manager-panel-trigger dropdown-block-link citation-manager-trigger-target" href="#">Citation manager</a></li></ul></div></div></div>
|
||
</div>
|
||
|
||
|
||
|
||
<div class="display-options" data-format-key="format"
|
||
data-default-format-value="abstract"
|
||
data-current-format-value="abstract" role="region">
|
||
<button
|
||
class="trigger usa-button"
|
||
title="Change format"
|
||
data-ga-category="display_options"
|
||
data-ga-action="display_options_dialog">
|
||
<span class="button-label">Display options</span>
|
||
</button>
|
||
|
||
<div class="dropdown" hidden>
|
||
<div class="title">
|
||
Display options
|
||
</div>
|
||
|
||
<div class="content">
|
||
|
||
<div class="format-container">
|
||
<span class="option-label">
|
||
Format
|
||
</span>
|
||
<select name="format" data-initial-value="abstract" aria-label="Display format" id="article-display-format">
|
||
<option value="abstract">Abstract</option>
|
||
|
||
<option value="pubmed">PubMed</option>
|
||
|
||
<option value="pmid">PMID</option>
|
||
|
||
</select>
|
||
</div>
|
||
|
||
</div>
|
||
</div>
|
||
</div>
|
||
<input type="hidden" name="csrfmiddlewaretoken" value="S2uVCHDz52oIHUIQ2V4MwuKDuQA3h9Mh9Pe0fx6VFDbcBlOGFeHO0Cgh2UifaKkB">
|
||
</div>
|
||
</div>
|
||
</div>
|
||
|
||
<div id="article-panels-container" class="article-panels-container">
|
||
<div id="save-action-panel" class="save-action-panel action-panel" aria-hidden="true">
|
||
<div class="inner-wrap">
|
||
<h2 class="action-panel-heading">
|
||
Save citation to file
|
||
</h2>
|
||
<form id="save-action-panel-form"
|
||
class="action-panel-content action-form"
|
||
action="/results-export-ids/"
|
||
data-by-search-action="/results-export-search-data/"
|
||
data-by-ids-action="/results-export-ids/"
|
||
method="post"
|
||
data-by-search-method="post"
|
||
data-by-ids-method="post">
|
||
|
||
<input type="hidden" name="csrfmiddlewaretoken" value="S2uVCHDz52oIHUIQ2V4MwuKDuQA3h9Mh9Pe0fx6VFDbcBlOGFeHO0Cgh2UifaKkB">
|
||
|
||
|
||
|
||
<div class="action-panel-control-wrap">
|
||
<label for="save-action-format" class="action-panel-label">
|
||
Format:
|
||
</label>
|
||
<select id="save-action-format" name="results-format" class="action-panel-selector">
|
||
<option value="summary-text">Summary (text)</option>
|
||
<option value="pubmed-text">PubMed</option>
|
||
<option value="pmid">PMID</option>
|
||
<option value="abstract">Abstract (text)</option>
|
||
<option value="csv">CSV</option>
|
||
</select>
|
||
</div>
|
||
|
||
<div class="action-panel-actions">
|
||
<button class="action-panel-submit"
|
||
type="submit"
|
||
data-loading-label="Saving..."
|
||
data-ga-category="save_share"
|
||
data-ga-action="save"
|
||
data-ga-label="save">
|
||
Create file
|
||
</button>
|
||
<button class="action-panel-cancel"
|
||
aria-label="Close 'Save citations to file' panel"
|
||
ref="linksrc=close_save_panel"
|
||
aria-controls="save-action-panel"
|
||
aria-expanded="false"
|
||
data-ga-category="save_share"
|
||
data-ga-action="save"
|
||
data-ga-label="cancel">
|
||
Cancel
|
||
</button>
|
||
</div>
|
||
</form>
|
||
</div>
|
||
</div>
|
||
|
||
|
||
|
||
|
||
<div id="email-action-panel"
|
||
class="email-action-panel action-panel"
|
||
aria-hidden="true"
|
||
data-recaptcha-script-url="https://www.recaptcha.net/recaptcha/enterprise.js?render=explicit&onload=onloadCaptchaCallback"
|
||
data-recaptcha-enabled="true">
|
||
<div class="inner-wrap">
|
||
<h2 class="action-panel-heading">
|
||
Email citation
|
||
</h2>
|
||
<form id="email-action-panel-form"
|
||
class="action-panel-content action-form"
|
||
action="/send-email/"
|
||
data-by-search-action="/results-export-email-by-search-data/"
|
||
data-by-ids-action="/send-email/"
|
||
data-by-search-method="post"
|
||
data-by-ids-method="post"
|
||
method="post"
|
||
>
|
||
<input type="hidden" name="csrfmiddlewaretoken" value="S2uVCHDz52oIHUIQ2V4MwuKDuQA3h9Mh9Pe0fx6VFDbcBlOGFeHO0Cgh2UifaKkB">
|
||
|
||
<div class="action-panel-control-wrap">
|
||
<span class="action-panel-label">
|
||
Subject:
|
||
</span>
|
||
<span class="email-subject" id="email-subject" aria-label="Email subject">1 selected item: 28031477 - PubMed</span>
|
||
<input type="hidden" name="email-subject" value="1 selected item: 28031477"/>
|
||
<input type="hidden" name="email-subject-hash" value="6e778567e8d98d8ec0030e7a42aa31c848a59a6bb4fe8e0fce8b229597260bdd"/>
|
||
</div>
|
||
|
||
<div class="action-panel-control-wrap">
|
||
<label for="email-to" class="action-panel-label required-field-asterisk">
|
||
To:
|
||
</label>
|
||
<input type="email" aria-label="Recipient Email Address" placeholder="email@example.com" id="email-to" class="email-to" pattern="^[a-zA-Z0-9_.+-]+@[a-zA-Z0-9-]+\.[a-zA-Z0-9-.]+$" maxlength="100" required>
|
||
</div>
|
||
|
||
<div class="action-panel-control-wrap">
|
||
<label for="email-from" class="action-panel-label">
|
||
From:
|
||
</label>
|
||
<input type="email" aria-label="Sender Email Address" placeholder="email@example.com" id="email-from" class="email-from" pattern="^[a-zA-Z0-9_.+-]+@[a-zA-Z0-9-]+\.[a-zA-Z0-9-.]+$" maxlength="256">
|
||
</div>
|
||
|
||
|
||
|
||
<div class="action-panel-control-wrap">
|
||
<label for="email-citation-format" class="action-panel-label">
|
||
Format:
|
||
</label>
|
||
<select id="email-citation-format" name="citation-format" class="action-panel-selector email-citation-format">
|
||
<option selected="selected" value="summary">Summary</option>
|
||
<option value="summary-text">Summary (text)</option>
|
||
<option value="abstract">Abstract</option>
|
||
<option value="abstract-text">Abstract (text)</option>
|
||
|
||
</select>
|
||
</div>
|
||
<div class="include-supplemental-container">
|
||
<input type="checkbox" aria-label="Include MeSH and other data" name="include-supplemental" id="email-include-supplemental" class="email-include-supplemental">
|
||
<label for="email-include-supplemental" class="email-include-supplemental-label">MeSH and other data</label>
|
||
</div>
|
||
|
||
|
||
|
||
<div class="form-field recaptcha ">
|
||
|
||
|
||
|
||
<div class="g-recaptcha" id="id-recaptcha" data-sitekey="6LfsWHMdAAAAAClKbtOpjQ2pMjgsGxvv7NdZW9uI"></div>
|
||
|
||
|
||
|
||
|
||
|
||
</div>
|
||
<div id="captcha-error-message" class="usa-input-error-message captcha-validation-message" role="alert"></div>
|
||
|
||
|
||
|
||
<div class="action-panel-actions">
|
||
<button class="action-panel-submit"
|
||
type="submit"
|
||
data-loading-label="Sending..."
|
||
data-ga-category="save_share"
|
||
data-ga-action="email"
|
||
data-ga-label="send">
|
||
Send email
|
||
</button>
|
||
<button class="action-panel-cancel"
|
||
aria-label="Close 'Email citations' panel"
|
||
ref="linksrc=close_email_panel"
|
||
aria-controls="email-action-panel"
|
||
aria-expanded="false"
|
||
data-ga-category="save_share"
|
||
data-ga-action="email"
|
||
data-ga-label="cancel">
|
||
Cancel
|
||
</button>
|
||
</div>
|
||
<input type="hidden" name="email-search-details" value="" />
|
||
<input type="hidden" name="email-search-details-hash" value="0e42663a6c3bd85498fcb88798998fed7bfdc45d457db35281e41afe13cc0524" />
|
||
</form>
|
||
</div>
|
||
</div>
|
||
|
||
<div id="collections-action-panel"
|
||
class="collections-action-panel action-panel in-progress-dots-panel"
|
||
aria-hidden="true"
|
||
data-collections-open-panel-enabled="false"
|
||
data-collections-open-panel-url-hash="#open-collections-panel">
|
||
<div class="inner-wrap">
|
||
<h3 class="action-panel-heading">
|
||
Add to Collections
|
||
</h3>
|
||
|
||
|
||
<form id="collections-action-panel-form"
|
||
class="collections-action-panel-form action-panel-content action-form action-panel-smaller-selectors"
|
||
data-existing-collections-url="/list-existing-collections/"
|
||
data-add-to-existing-collection-url="/add-to-existing-collection/"
|
||
data-create-and-add-to-new-collection-url="/create-and-add-to-new-collection/"
|
||
data-get-article-ids-by-search-url="/get-article-ids-by-search/"
|
||
data-myncbi-max-collection-name-length="100"
|
||
data-add-to-collection-max-amount="1000"
|
||
data-collections-root-url="https://www.ncbi.nlm.nih.gov/myncbi/collections/">
|
||
|
||
<input type="hidden" name="csrfmiddlewaretoken" value="S2uVCHDz52oIHUIQ2V4MwuKDuQA3h9Mh9Pe0fx6VFDbcBlOGFeHO0Cgh2UifaKkB">
|
||
|
||
|
||
|
||
<div class="choice-group" role="radiogroup">
|
||
<ul class="radio-group-items">
|
||
<li>
|
||
<input type="radio"
|
||
id="collections-action-panel-new"
|
||
class="collections-new"
|
||
name="collections"
|
||
value="new"
|
||
data-ga-category="save_share"
|
||
data-ga-action="collections"
|
||
data-ga-label="collections_radio_new">
|
||
<label for="collections-action-panel-new">Create a new collection</label>
|
||
</li>
|
||
<li>
|
||
<input type="radio"
|
||
id="collections-action-panel-existing"
|
||
class="collections-existing"
|
||
name="collections"
|
||
value="existing"
|
||
checked="true"
|
||
data-ga-category="save_share"
|
||
data-ga-action="collections"
|
||
data-ga-label="collections_radio_existing">
|
||
<label for="collections-action-panel-existing">Add to an existing collection</label>
|
||
</li>
|
||
</ul>
|
||
</div>
|
||
|
||
<div class="controls-wrapper">
|
||
<div class="action-panel-control-wrap new-collections-controls">
|
||
<label for="collections-action-panel-add-to-new" class="action-panel-label required-field-asterisk">
|
||
Name your collection:
|
||
</label>
|
||
<input
|
||
type="text"
|
||
name="add-to-new-collection"
|
||
id="collections-action-panel-add-to-new"
|
||
class="collections-action-add-to-new"
|
||
pattern="[^"&=<>\/]*" title="The following characters are not allowed in the Name field: "&=<>/"
|
||
maxlength="100"
|
||
data-ga-category="save_share"
|
||
data-ga-action="create_collection"
|
||
data-ga-label="non_favorties_collection">
|
||
<div class="collections-new-name-too-long usa-input-error-message selection-validation-message">
|
||
Name must be less than 100 characters
|
||
</div>
|
||
</div>
|
||
<div class="action-panel-control-wrap existing-collections-controls">
|
||
<label for="collections-action-panel-add-to-existing" class="action-panel-label">
|
||
Choose a collection:
|
||
</label>
|
||
<select id="collections-action-panel-add-to-existing"
|
||
class="action-panel-selector collections-action-add-to-existing"
|
||
data-ga-category="save_share"
|
||
data-ga-action="select_collection"
|
||
data-ga-label="($('#collections-action-add-to-existing').val() === 'Favorites') ? 'Favorites' : 'non_favorites_collection'">
|
||
</select>
|
||
<div class="collections-retry-load-on-error usa-input-error-message selection-validation-message">
|
||
Unable to load your collection due to an error<br>
|
||
<a href="#">Please try again</a>
|
||
</div>
|
||
</div>
|
||
</div>
|
||
|
||
<div class="action-panel-actions">
|
||
<button class="action-panel-submit"
|
||
type="submit"
|
||
data-loading-label="Adding..."
|
||
data-pinger-ignore
|
||
data-ga-category="save_share"
|
||
data-ga-action="collections"
|
||
data-ga-label="add">
|
||
Add
|
||
</button>
|
||
<button class="action-panel-cancel"
|
||
aria-label="Close 'Add to Collections' panel"
|
||
ref="linksrc=close_collections_panel"
|
||
aria-controls="collections-action-panel"
|
||
aria-expanded="false"
|
||
data-ga-category="save_share"
|
||
data-ga-action="collections"
|
||
data-ga-label="cancel">
|
||
Cancel
|
||
</button>
|
||
</div>
|
||
</form>
|
||
</div>
|
||
</div>
|
||
|
||
<div id="bibliography-action-panel"
|
||
class="bibliography-action-panel action-panel in-progress-dots-panel"
|
||
aria-hidden="true"
|
||
data-bibliography-open-panel-enabled="false"
|
||
data-bibliography-open-panel-url-hash="#open-bibliography-panel">
|
||
<div class="inner-wrap">
|
||
<h3 class="action-panel-heading">
|
||
Add to My Bibliography
|
||
</h3>
|
||
<form id="bibliography-action-panel-form"
|
||
class="bibliography-action-panel-form action-panel-content action-form action-panel-smaller-selectors"
|
||
data-add-to-bibliography-max-amount="100"
|
||
data-add-to-bibliography-batch-size="10"
|
||
data-bibliography-delegates-url="/list-bibliography-delegates/"
|
||
data-add-to-bibliography-url="/add-to-bibliography/"
|
||
data-get-article-ids-by-search-url="/get-article-ids-by-search/"
|
||
data-mybib-root-url="https://www.ncbi.nlm.nih.gov/myncbi/collections/mybibliography/">
|
||
|
||
<input type="hidden" name="csrfmiddlewaretoken" value="S2uVCHDz52oIHUIQ2V4MwuKDuQA3h9Mh9Pe0fx6VFDbcBlOGFeHO0Cgh2UifaKkB">
|
||
|
||
|
||
|
||
<div class="action-panel-control-wrap bibliographies-controls">
|
||
<div class="choice-group">
|
||
<ul class="bibliographies-action-add radio-group-items">
|
||
<li>
|
||
<input name="bibliography" id="my-bibliography" class="my-bibliography" type="radio" checked/>
|
||
<label for="my-bibliography">My Bibliography</label>
|
||
</li>
|
||
</ul>
|
||
</div>
|
||
</div>
|
||
|
||
<div class="bibliographies-retry-load-on-error usa-input-error-message selection-validation-message">
|
||
Unable to load your delegates due to an error<br>
|
||
<a href="#">Please try again</a>
|
||
</div>
|
||
|
||
<div class="action-panel-actions">
|
||
<button class="action-panel-submit"
|
||
type="submit"
|
||
data-loading-label="Adding..."
|
||
data-pinger-ignore>
|
||
Add
|
||
</button>
|
||
<button class="action-panel-cancel"
|
||
aria-label="Close 'Add to bibliography' panel"
|
||
ref="linksrc=close_bibliography_panel"
|
||
aria-controls="bibliography-action-panel"
|
||
aria-expanded="false"
|
||
data-ga-category="save_share"
|
||
data-ga-action="mybib"
|
||
data-ga-label="cancel">
|
||
Cancel
|
||
</button>
|
||
</div>
|
||
</form>
|
||
</div>
|
||
</div>
|
||
|
||
|
||
|
||
|
||
<div id="saved-search-action-panel" class="saved-search-action-panel action-panel " aria-hidden="true"
|
||
data-saved-search-open-panel-enabled="false"
|
||
data-saved-search-open-panel-url-hash="#open-saved-search-panel">
|
||
<div class="inner-wrap">
|
||
<h2 class="action-panel-heading">
|
||
Your saved search
|
||
</h2>
|
||
|
||
<form id="saved-search-action-panel-form"
|
||
class="saved-search-action-panel-form action-panel-content action-form"
|
||
data-create-saved-search-url="/create-saved-search/"
|
||
data-try-search-terms-url="/try-search-term/"
|
||
data-saved-search-root-url="https://www.ncbi.nlm.nih.gov/myncbi/searches/">
|
||
|
||
<input type="hidden" name="csrfmiddlewaretoken" value="S2uVCHDz52oIHUIQ2V4MwuKDuQA3h9Mh9Pe0fx6VFDbcBlOGFeHO0Cgh2UifaKkB">
|
||
|
||
<div class="action-panel-control-wrap">
|
||
<label for="saved-search-name" class="action-panel-label saved-search-name-label required-field-asterisk">
|
||
Name of saved search:
|
||
</label>
|
||
<input maxlength="200"
|
||
type="text"
|
||
name="saved-search-name"
|
||
id="saved-search-name"
|
||
class="saved-search-name"
|
||
value=""
|
||
required
|
||
pattern="[^"&=<>\/]*" title="The following characters are not allowed in the Name field: "&=<>/">
|
||
</div>
|
||
|
||
<div class="action-panel-control-wrap">
|
||
<label for="saved-search-term" class="action-panel-label required-field-asterisk">
|
||
Search terms:
|
||
</label>
|
||
<textarea name="saved-search-term" id="saved-search-term" class="saved-search-term" required></textarea>
|
||
</div>
|
||
<div class="test-search-term-wrap">
|
||
<a href="#" class="try-search-term">Test search terms</a>
|
||
</div>
|
||
<div class="choice-group action-panel-extra-margin-top">
|
||
<span class="action-panel-label" id="fieldset-label">
|
||
Would you like email updates of new search results?
|
||
</span>
|
||
<fieldset id="saved-search-alert" aria-describedby="fieldset-label">
|
||
<legend class="usa-sr-only">Saved Search Alert Radio Buttons</legend>
|
||
<ul class="radio-group-items">
|
||
<li>
|
||
<input type="radio" id="saved-search-alert-yes" class="saved-search-alert-yes" name="saved-search-alert" value="yes" checked>
|
||
<label for="saved-search-alert-yes" class="action-panel-label">Yes</label>
|
||
</li>
|
||
<li>
|
||
<input aria-label="No radio input" type="radio" id="saved-search-alert-no" class="saved-search-alert-no" name="saved-search-alert" value="no">
|
||
<label for="saved-search-alert-no" class="action-panel-label">No</label>
|
||
</li>
|
||
</ul>
|
||
</fieldset>
|
||
</div>
|
||
|
||
<div class="alert-schedule-wrap">
|
||
<div class="action-panel-control-wrap">
|
||
<label class="action-panel-label">
|
||
Email:
|
||
</label>
|
||
<span aria-label="Email address" id="saved-search-email" class="action-panel-label"><span class="action-panel-label-bold"></span> (<a class="myncbi-account-settings" href="https://www.ncbi.nlm.nih.gov/account/settings/">change</a>)</span>
|
||
</div>
|
||
<div class="action-panel-control-wrap action-panel-extra-margin-top">
|
||
<label for="saved-search-frequency" class="action-panel-label">
|
||
Frequency:
|
||
</label>
|
||
<select id="saved-search-frequency" class="no-border-panel-selector saved-search-frequency">
|
||
<option value="monthly">Monthly</option>
|
||
<option value="weekly">Weekly</option>
|
||
<option value="daily">Daily</option>
|
||
</select>
|
||
</div>
|
||
<div class="action-panel-control-wrap saved-search-monthly-additional">
|
||
<label for="saved-search-monthly-on-day" class="action-panel-label">
|
||
Which day?
|
||
</label>
|
||
<select id="saved-search-monthly-on-day" class="no-border-panel-selector">
|
||
<option value="Sunday">The first Sunday</option>
|
||
<option value="Monday">The first Monday</option>
|
||
<option value="Tuesday">The first Tuesday</option>
|
||
<option value="Wednesday">The first Wednesday</option>
|
||
<option value="Thursday">The first Thursday</option>
|
||
<option value="Friday">The first Friday</option>
|
||
<option value="Saturday">The first Saturday</option>
|
||
<option value="day">The first day</option>
|
||
<option value="weekday">The first weekday</option>
|
||
</select>
|
||
</div>
|
||
<div class="action-panel-control-wrap saved-search-weekly-additional">
|
||
<label for="saved-search-weekly-on-day" class="action-panel-label">
|
||
Which day?
|
||
</label>
|
||
<select id="saved-search-weekly-on-day" class="no-border-panel-selector saved-search-weekly-on-day">
|
||
<option value="Sunday">Sunday</option>
|
||
<option value="Monday">Monday</option>
|
||
<option value="Tuesday">Tuesday</option>
|
||
<option value="Wednesday">Wednesday</option>
|
||
<option value="Thursday">Thursday</option>
|
||
<option value="Friday">Friday</option>
|
||
<option value="Saturday">Saturday</option>
|
||
</select>
|
||
</div>
|
||
<div class="action-panel-control-wrap">
|
||
<label for="saved-search-report" class="action-panel-label">
|
||
Report format:
|
||
</label>
|
||
<select id="saved-search-report" class="no-border-panel-selector saved-search-report">
|
||
<option value="DocSum">Summary</option>
|
||
<option value="DocSumText">Summary (text)</option>
|
||
<option value="Abstract">Abstract</option>
|
||
<option value="AbstractText">Abstract (text)</option>
|
||
<option value="MEDLINE">PubMed</option>
|
||
</select>
|
||
</div>
|
||
<div class="action-panel-control-wrap">
|
||
<label for="saved-search-amount" class="action-panel-label">
|
||
Send at most:
|
||
</label>
|
||
<select id="saved-search-amount" class="no-border-panel-selector saved-search-amount">
|
||
<option value="1">1 item</option>
|
||
<option value="5" selected>5 items</option>
|
||
<option value="10">10 items</option>
|
||
<option value="20">20 items</option>
|
||
<option value="50">50 items</option>
|
||
<option value="100">100 items</option>
|
||
<option value="200">200 items</option>
|
||
</select>
|
||
</div>
|
||
<div>
|
||
<input type="checkbox" id="saved-search-send-if-no-result" class="saved-search-send-if-no-result" name="saved-search-send-if-no-result">
|
||
<label for="saved-search-send-if-no-result" class="action-panel-label smaller-checkbox">
|
||
Send even when there aren't any new results
|
||
</label>
|
||
</div>
|
||
<div class="action-panel-control-wrap option-text-in-email-wrap">
|
||
<label for="saved-search-email-text" class="action-panel-label">
|
||
Optional text in email:
|
||
</label>
|
||
<textarea name="saved-search-email-text"
|
||
id="saved-search-email-text"
|
||
class="saved-search-email-text"></textarea>
|
||
</div>
|
||
|
||
</div>
|
||
|
||
<div class="action-panel-actions">
|
||
<button class="action-panel-submit"
|
||
type="submit"
|
||
data-loading-label="Saving..."
|
||
data-ga-category="save_share"
|
||
data-ga-action="alert"
|
||
data-ga-label="save">
|
||
Save
|
||
</button>
|
||
<button class="action-panel-cancel"
|
||
aria-label="Close 'Your saved search' panel"
|
||
ref="linksrc=close_saved_search_panel"
|
||
aria-controls="saved-search-action-panel"
|
||
aria-expanded="false"
|
||
data-ga-category="save_share"
|
||
data-ga-action="alert"
|
||
data-ga-label="cancel">
|
||
Cancel
|
||
</button>
|
||
</div>
|
||
</form>
|
||
</div>
|
||
</div>
|
||
|
||
<div id="citation-manager-action-panel" class="citation-manager-action-panel action-panel" aria-hidden="true">
|
||
<div class="inner-wrap">
|
||
<h2 class="action-panel-heading">
|
||
Create a file for external citation management software
|
||
</h2>
|
||
<form id="citation-manager-action-panel-form"
|
||
class="action-panel-content action-form"
|
||
action="/results-export-ids/"
|
||
data-by-search-action="/results-export-search-data/"
|
||
data-by-ids-action="/results-export-ids/"
|
||
method="post"
|
||
data-by-search-method="post"
|
||
data-by-ids-method="post">
|
||
|
||
<input type="hidden" name="csrfmiddlewaretoken" value="S2uVCHDz52oIHUIQ2V4MwuKDuQA3h9Mh9Pe0fx6VFDbcBlOGFeHO0Cgh2UifaKkB">
|
||
|
||
|
||
|
||
<input name="results-format" type="hidden" value="pubmed"/>
|
||
|
||
<div class="action-panel-actions">
|
||
<button class="action-panel-submit"
|
||
type="submit"
|
||
data-loading-label="Sending..."
|
||
data-ga-category="save_share"
|
||
data-ga-action="citation_manager"
|
||
data-ga-label="save">
|
||
Create file
|
||
</button>
|
||
<button class="action-panel-cancel"
|
||
aria-label="Close 'Send citations to citation manager' panel"
|
||
ref="linksrc=close_citation_manager_panel"
|
||
aria-controls="citation-manager-action-panel"
|
||
aria-expanded="false"
|
||
data-ga-category="save_share"
|
||
data-ga-action="citation_manager"
|
||
data-ga-label="cancel">
|
||
Cancel
|
||
</button>
|
||
</div>
|
||
</form>
|
||
</div>
|
||
</div>
|
||
|
||
|
||
|
||
|
||
<div id="rss-action-panel" class="rss-action-panel action-panel " aria-hidden="true">
|
||
<div class="inner-wrap">
|
||
<h2 class="action-panel-heading">
|
||
Your RSS Feed
|
||
</h2>
|
||
|
||
<form id="rss-action-panel-form"
|
||
class="rss-action-panel-form action-panel-content action-form"
|
||
data-create-rss-feed-url="/create-rss-feed-url/"
|
||
data-search-form-term-value="">
|
||
|
||
<input type="hidden" name="csrfmiddlewaretoken" value="S2uVCHDz52oIHUIQ2V4MwuKDuQA3h9Mh9Pe0fx6VFDbcBlOGFeHO0Cgh2UifaKkB">
|
||
|
||
<div class="action-panel-control-wrap">
|
||
<label for="rss-name" class="action-panel-label required-field-asterisk">
|
||
Name of RSS Feed:
|
||
</label>
|
||
<input maxlength="200"
|
||
placeholder="Name"
|
||
type="text"
|
||
name="rss-name"
|
||
id="rss-name"
|
||
class="rss-name"
|
||
value=''
|
||
required
|
||
pattern="[^"&=<>\/]*" title="The following characters are not allowed in the Name field: "&=<>/">
|
||
</div>
|
||
|
||
<div class="rss-limit-wrap">
|
||
<div class="action-panel-control-wrap action-panel-extra-margin-top">
|
||
<label for="rss-limit" class="action-panel-label">
|
||
Number of items displayed:
|
||
</label>
|
||
<select id="rss-limit" class="no-border-panel-selector rss-limit">
|
||
<option value="5">5</option>
|
||
<option value="10">10</option>
|
||
<option value="15" selected="selected">15</option>
|
||
<option value="20">20</option>
|
||
<option value="50">50</option>
|
||
<option value="100">100</option>
|
||
</select>
|
||
</div>
|
||
</div>
|
||
|
||
<div class="action-panel-actions">
|
||
<button class="action-panel-submit"
|
||
type="submit"
|
||
data-loading-label="Creating..."
|
||
data-ga-category="save_share"
|
||
data-ga-action="alert"
|
||
data-ga-label="save">
|
||
Create RSS
|
||
</button>
|
||
<button class="action-panel-cancel"
|
||
aria-label="Close 'Your RSS' panel"
|
||
ref="linksrc=close_rss_panel"
|
||
aria-controls="rss-action-panel"
|
||
aria-expanded="false"
|
||
data-ga-category="save_share"
|
||
data-ga-action="alert"
|
||
data-ga-label="cancel">
|
||
Cancel
|
||
</button>
|
||
</div>
|
||
|
||
<div class="action-panel-control-wrap rss-link-copy-wrap">
|
||
<label for="rss-link" class="usa-sr-only">RSS Link</label>
|
||
<input placeholder="Your RSS Feed Link" type="text" name="rss-link" id="rss-link" class="rss-link" title="RSS Link">
|
||
<button
|
||
type="button"
|
||
disabled
|
||
class="rss-link-copy-button disabled"
|
||
data-ga-category="save_share"
|
||
data-ga-action="rss"
|
||
data-ga-label="copy">
|
||
Copy
|
||
</button>
|
||
</div>
|
||
</form>
|
||
</div>
|
||
</div>
|
||
|
||
</div>
|
||
</div>
|
||
|
||
<div class="article-page" id="article-page" data-article-pmid="28031477">
|
||
<aside class="page-sidebar">
|
||
<div class="inner-wrap">
|
||
|
||
<div class="full-text-links">
|
||
<div class="full-view">
|
||
<h3 class="title">
|
||
Full text links
|
||
</h3>
|
||
<div class="full-text-links-list">
|
||
<a class="link-item
|
||
|
||
dialog-focus"
|
||
href="https://rupress.org/jem/article-lookup/doi/10.1084/jem.20161452"
|
||
target="_blank"
|
||
rel="noopener"
|
||
|
||
ref="linksrc=fulltextorjournal_fulltext&is_pmc=False&PrId=7898&itool=Abstract-def&log$=linkouticon&uid=28031477&db=pubmed&nlmid=2985109R"
|
||
title="See full text options at Silverchair Information Systems"
|
||
|
||
data-ga-category="full_text"
|
||
data-ga-action="Silverchair Information Systems"
|
||
data-ga-label="28031477"
|
||
><img src="https://cdn.ncbi.nlm.nih.gov/corehtml/query/egifs/https:--rupress.org-images-pubmed-jem_free.png" alt="Silverchair Information Systems full text link"><span class="text">
|
||
|
||
Silverchair Information Systems
|
||
|
||
</span></a><a class="link-item
|
||
pmc
|
||
"
|
||
href="https://pmc.ncbi.nlm.nih.gov/articles/pmid/28031477/"
|
||
target="_blank"
|
||
rel="noopener"
|
||
|
||
ref="linksrc=fulltextorjournal_fulltext&is_pmc=True&PrId=3494&itool=Abstract-def&log$=linkouticon&uid=28031477&db=pubmed&nlmid=2985109R"
|
||
title="Free full text at PubMed Central"
|
||
|
||
data-ga-category="full_text"
|
||
data-ga-action="PMC"
|
||
data-ga-label="28031477"
|
||
><span class="text">
|
||
|
||
Free PMC article
|
||
|
||
</span></a>
|
||
</div>
|
||
|
||
</div>
|
||
|
||
<div class="short-view">
|
||
<a href="#" class="full-text-links-button full-text-links-dialog-trigger">
|
||
Full text links
|
||
</a>
|
||
</div>
|
||
</div>
|
||
|
||
|
||
|
||
|
||
<div class="actions-buttons sidebar"><h3 class="title">Actions</h3><div class="inner-wrap"><button class="citation-button citation-dialog-trigger"
|
||
aria-label="Open dialog with citation text in different styles"
|
||
data-ga-category="save_share"
|
||
data-ga-action="cite"
|
||
data-ga-label="open"
|
||
data-all-citations-url="/28031477/citations/"
|
||
data-citation-style="nlm"
|
||
data-pubmed-format-link="/28031477/export/"><span class="button-label">Cite</span></button><link type="text/css" href="ncbi-overlay-block/src/overlay-block.css"><div class="collections-button-container" data-article-id="28031477" data-article-db="pubmed"><button class="collections-button collections-dialog-trigger"
|
||
aria-label="Save article in MyNCBI collections."
|
||
data-ga-category="collections_button"
|
||
data-ga-action="click"
|
||
data-ga-label="collections_button"
|
||
data-collections-open-dialog-enabled="false"
|
||
data-collections-open-dialog-url="https://account.ncbi.nlm.nih.gov/?back_url=https%3A%2F%2Fpubmed.ncbi.nlm.nih.gov%2F28031477%2F%23open-collections-dialog"
|
||
data-in-collections="false"><span class="button-label">Collections</span></button><div class="overlay" role="dialog"><div id="collections-action-dialog"
|
||
class="dialog collections-dialog"
|
||
aria-hidden="true"><div class="title">Add to Collections</div><div class="collections-action-panel action-panel"><form id="collections-action-dialog-form"
|
||
class="collections-action-panel-form action-panel-content action-form action-panel-smaller-selectors"
|
||
data-existing-collections-url="/list-existing-collections/"
|
||
data-add-to-existing-collection-url="/add-to-existing-collection/"
|
||
data-create-and-add-to-new-collection-url="/create-and-add-to-new-collection/"
|
||
data-myncbi-max-collection-name-length="100"
|
||
data-collections-root-url="https://www.ncbi.nlm.nih.gov/myncbi/collections/"><input type="hidden" name="csrfmiddlewaretoken" value="S2uVCHDz52oIHUIQ2V4MwuKDuQA3h9Mh9Pe0fx6VFDbcBlOGFeHO0Cgh2UifaKkB"><div class="choice-group" role="radiogroup"><ul class="radio-group-items"><li><input type="radio"
|
||
id="collections-action-dialog-new"
|
||
class="collections-new"
|
||
name="collections"
|
||
value="new"
|
||
data-ga-category="collections_button"
|
||
data-ga-action="click"
|
||
data-ga-label="collections_radio_new"><label for="collections-action-dialog-new">Create a new collection</label></li><li><input type="radio"
|
||
id="collections-action-dialog-existing"
|
||
class="collections-existing"
|
||
name="collections"
|
||
value="existing"
|
||
checked="true"
|
||
data-ga-category="collections_button"
|
||
data-ga-action="click"
|
||
data-ga-label="collections_radio_existing"><label for="collections-action-dialog-existing">Add to an existing collection</label></li></ul></div><div class="controls-wrapper"><div class="action-panel-control-wrap new-collections-controls"><label for="collections-action-dialog-add-to-new" class="action-panel-label required-field-asterisk">
|
||
Name your collection:
|
||
</label><input
|
||
type="text"
|
||
name="add-to-new-collection"
|
||
id="collections-action-dialog-add-to-new"
|
||
class="collections-action-add-to-new"
|
||
pattern="[^"&=<>\/]*" title="The following characters are not allowed in the Name field: "&=<>/"
|
||
maxlength="100"
|
||
data-ga-category="collections_button"
|
||
data-ga-action="create_collection"
|
||
data-ga-label="non_favorties_collection"><div class="collections-new-name-too-long usa-input-error-message selection-validation-message">
|
||
Name must be less than 100 characters
|
||
</div></div><div class="action-panel-control-wrap existing-collections-controls"><label for="collections-action-dialog-add-to-existing" class="action-panel-label">
|
||
Choose a collection:
|
||
</label><select id="collections-action-dialog-add-to-existing"
|
||
class="action-panel-selector collections-action-add-to-existing"
|
||
data-ga-category="collections_button"
|
||
data-ga-action="select_collection"
|
||
data-ga-label="($('.collections-action-add-to-existing').val() === 'Favorites') ? 'Favorites' : 'non_favorites_collection'"></select><div class="collections-retry-load-on-error usa-input-error-message selection-validation-message">
|
||
Unable to load your collection due to an error<br><a href="#">Please try again</a></div></div></div><div class="action-panel-actions"><button class="action-panel-submit"
|
||
type="submit"
|
||
data-loading-label="Adding..."
|
||
data-pinger-ignore
|
||
data-ga-category="collections_button"
|
||
data-ga-action="click"
|
||
data-ga-label="add">
|
||
Add
|
||
</button><button class="action-panel-cancel"
|
||
aria-label="Close 'Add to Collections' panel"
|
||
ref="linksrc=close_collections_panel"
|
||
aria-controls="collections-action-panel"
|
||
aria-expanded="false"
|
||
data-ga-category="collections_button"
|
||
data-ga-action="click"
|
||
data-ga-label="cancel">
|
||
Cancel
|
||
</button></div></form></div></div></div></div><button class="more-actions-button more-actions-dialog-trigger"
|
||
title="Open dialog with more actions to take"
|
||
ref="linksrc=more_actions_btn"></button><div class="display-options" data-format-key="format"
|
||
data-default-format-value="abstract"
|
||
data-current-format-value="abstract" role="region"><button
|
||
class="trigger usa-button"
|
||
title="Change format"
|
||
data-ga-category="display_options"
|
||
data-ga-action="display_options_dialog"><span class="button-label">Display options</span></button><div class="dropdown" hidden><div class="title">
|
||
Display options
|
||
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Lipin-2 regulates NLRP3 inflammasome by affecting P2X7 receptor activation
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5
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Lipin-2 regulates NLRP3 inflammasome by affecting P2X7 receptor activation
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><sup class="key">5</sup> Instituto de Biología y Genética Molecular, Consejo Superior de Investigaciones Científicas, Universidad de Valladolid, 47003 Valladolid, Spain mbalboa@ibgm.uva.es.</li>
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Mutations in human LPIN2 produce a disease known as Majeed syndrome, the clinical manifestations of which are ameliorated by strategies that block IL-1β or its receptor. However the role of lipin-2 during IL-1β production remains elusive. We show here that lipin-2 controls excessive IL-1β formation in primary human and mouse macrophages by several mechanisms, including activation of the inflammasome NLRP3. Lipin-2 regulates MAPK activation, which mediates synthesis of pro-IL-1β during inflammasome priming. Lipin-2 also inhibits the activation and sensitization of the purinergic receptor P2X7 and K<sup>+</sup> efflux, apoptosis-associated speck-like protein with a CARD domain oligomerization, and caspase-1 processing, key events during inflammasome activation. Reduced levels of lipin-2 in macrophages lead to a decrease in cellular cholesterol levels. In fact, restoration of cholesterol concentrations in cells lacking lipin-2 decreases ion currents through the P2X7 receptor, and downstream events that drive IL-1β production. Furthermore, lipin-2-deficient mice exhibit increased sensitivity to high lipopolysaccharide doses. Collectively, our results unveil lipin-2 as a critical player in the negative regulation of NLRP3 inflammasome.
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© 2017 Lordén et al.
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<p> Figure 1. </p>
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<div class="figure-caption-contents"><b> The effect of lipin-2 on… </b></div>
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<div class="figure-caption-contents"><b> The effect of lipin-2 on IL-1β production by murine and human macrophages. </b> IL-1β…</div>
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<div class="figure-caption-contents"><b>The effect of lipin-2 on IL-1β production by murine and human macrophages.</b> IL-1β presence in supernatants from primary human macrophages (A), BMDMs from WT and <i>Lpin2<sup>−/−</sup></i> mice (B and D), or RAW264.7 cells (C) stimulated with 200 ng/ml LPS for 4 h, 2 mM ATP for 40 min, or both, as indicated, were quantified by specific ELISAs. (A–C) Human macrophages and RAW264.7 cells were silenced as described in Materials and methods, using control (siRNACtrl.) and lipin-2–specific siRNAs (siRNA<i>Lpin2</i>). (right) <i>Lpin2</i> mRNA levels at the time of stimulation. (D) BMDMs supernatants were analyzed by immunoblot using specific antibodies against IL-1β. (right) Densitometric quantification of the bands. (E) IL-1β production was analyzed in BMDMs supernatants from WT, <i>Nlrp3</i><sup>−/−</sup>, <i>Asc</i><sup>−/−</sup>, and <i>Casp1</i><sup>−/−</sup> animals treated with a control siRNA (siRNACtrl.) or siRNAs against <i>Lpin2</i> (siRNA<i>Lpin2</i>) for 2 d, and then stimulated as indicated. (right) Level of <i>Lpin2</i> mRNA after silencing. Data from A–C and E are shown as means ± SD and are representative of at least three independent experiments made in triplicate. Data from D are representative of at least three independent experiments. *, P < 0.05; **, P < 0.01; ***, P < 0.001, Student’s <i>t</i> test.</div>
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<div class="figure-caption-contents"><b> The effect of lipin-2 on TLR4 signaling. </b> (A) WT and <i> Lpin2 <sup> −/− </sup> </i> BMDMs…</div>
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<div class="figure-caption-contents"><b>The effect of lipin-2 on TLR4 signaling.</b> (A) WT and <i>Lpin2<sup>−/−</sup></i> BMDMs were stimulated with 200 ng/ml LPS for the indicated periods of time, and then mRNA levels for <i>Tnf</i> were quantified by RT-qPCR. (B) Quantification of TNF present in cellular supernatants from WT and <i>Lpin2<sup>−/−</sup></i> BMDMs (left) or silenced human macrophages (right) treated with 200 ng/ml LPS for 4 h. (C) Quantification of <i>Il1b</i> mRNA levels from WT and <i>Lpin2<sup>−/−</sup></i> BMDMs (left), or silenced human macrophages (right) stimulated with LPS. (D) Analysis by Immunoblot of pro–IL-1β present in homogenates from BMDMs (left). (right) Relative expression levels of IL-1β against β-actin. (E) Analysis by immunoblot of MAPKs and their phosphorylated forms present in BMDMs activated as in A (left). (right) Relative quantification of phosphoproteins against total proteins. (F) Control (siRNACtrl.) and lipin-2–silenced (siRNALpin2) RAW264.7 cells were pretreated with 10 µM PD98059, SP600125, or SB203580 for 30 min, and then stimulated with 200 ng/ml LPS for 4 h. <i>Il1b</i> mRNA levels were quantified by RT-qPCR. (G) WT and <i>Lpin2<sup>−/−</sup></i> BMDMs were treated with inhibitors before or after stimulation with LPS. Stimulations were as indicated. IL-1β present in cellular supernatants is shown. (H) WT and <i>Lpin2<sup>−/−</sup></i> BMDMs were stimulated with LPS as in A, and then Nlrp3 mRNA levels were quantified. (I) Analysis by immunoblot of NLRP3 present in cellular homogenates from BMDMs stimulated as in A (left). (right) Relative NLRP3 expression levels against β actin. β-Actin bands are the same as shown in E because the same blot was used. Data from A–C and F–H are shown as mean ± SD, and experiments shown are representative of at least three independent experiments made in triplicate. Data from D, E, and I are representative of at least three independent experiments. *, P < 0.05; **, P < 0.01; ***, P < 0.001, Student’s <i>t</i> test. <sup>#</sup>, P < 0.05; <sup>###</sup>, P < 0.001, versus siRNALpin2 + LPS cells (F), or <i>Lpin2<sup>−/−</sup></i> + LPS/ATP cells (G) by Student’s <i>t</i> test.</div>
|
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<p> Figure 3. </p>
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<div class="figure-caption-contents"><b> The effect of lipin-2 on… </b></div>
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<p> Figure 3. </p>
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<div class="figure-caption-contents"><b> The effect of lipin-2 on caspase-1 activation. </b> (A) WT and <i> Lpin2 <sup> −/− </sup> </i> BMDMs…</div>
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|
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<div class="figure-caption-contents"><b>The effect of lipin-2 on caspase-1 activation.</b> (A) WT and <i>Lpin2<sup>−/−</sup></i> BMDMs (left) or silenced human macrophages (right) were stimulated with 200 ng/ml LPS for 4 h or the indicated periods of time, 2 mM ATP for 40 min, or both, as indicated. IL-18 present in cellular supernatants was quantified by specific ELISAs. (B) Cells as in A were stimulated with LPS and <i>Il18</i> mRNA levels were quantified by RT-qPCR. (C) Analysis by immunoblot of active caspase-1 present in supernatants from WT and <i>Lpin2<sup>−/−</sup></i> BMDMs (top). (bottom) Densitometric quantification of the bands. (D) Analysis of intracellular active caspase by flow cytometry in RAW264.7 cells treated with control siRNA (siRNACtrl.) or siRNA against lipin-2 (siRNA<i>Lpin2</i>) and stimulated as in A. Median fluorescence intensities (MFI) are indicated (left). (right) Percentage of cells with active caspase-1. (E and F) WT and <i>Lpin2<sup>−/−</sup></i> BMDMs (E) or silenced human macrophages (F) were pretreated with 10 µM YVAD or ZVAD for 30 min and then stimulated as in A. (G) IL-1β (left) and TNF (right) present in cellular supernatants were quantified by specific ELISAs. LDH release from RAW264.7 cells treated as in D. (H) Analysis by flow cytometry of RAW264.7 cells treated as in D and stained with propidium iodide (PI; left). (right) Percentage of cells positive for PI. Data from A, B, and E–G are shown as mean ± SD, and experiments shown are representative of at least three independent experiments made in triplicate. Data from C, D, and H are representative of at least three independent experiments. *, P < 0.05; **, P < 0.01; ***, P < 0.001, Student’s <i>t</i> test. (E) <sup>###</sup>, P < 0.001, versus <i>Lpin2</i><sup>−/−</sup> + LPS/ATP cells by Student’s <i>t</i> test.</div>
|
||
|
||
|
||
|
||
|
||
</figcaption>
|
||
|
||
|
||
</figure>
|
||
|
||
<figure class="figure-item "
|
||
itemscope itemtype="http://schema.org/ImageObject"
|
||
itemprop="associatedMedia"
|
||
data-slide-index="3"
|
||
data-label-slug="figure-4">
|
||
<a class="figure-link"
|
||
href="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ba50/5294860/6a109e4e05f4/JEM_20161452_Fig4.jpg"
|
||
itemprop="contentUrl"
|
||
aria-describedby="figure-caption-3"
|
||
role="button"
|
||
data-image-width="753"
|
||
data-image-height="758"
|
||
data-image-alt="Figure 4."
|
||
data-pmc-id="PMC5294860"
|
||
data-figure-id="fig4">
|
||
<img class="figure-thumb" itemprop="thumbnail"
|
||
id="article-image-3"
|
||
src="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ba50/5294860/c2628121114a/JEM_20161452_Fig4.gif"
|
||
alt="Figure 4." />
|
||
</a>
|
||
|
||
<meta itemprop="width" itemtype="http://schema.org/ImageObject" content="753">
|
||
<meta itemprop="height" itemtype="http://schema.org/ImageObject" content="758">
|
||
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||
|
||
<div class="figure-caption figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="caption" aria-hidden="true">
|
||
|
||
<div class="caption-wrap">
|
||
|
||
|
||
|
||
|
||
|
||
<strong class="figure-label">
|
||
<p> Figure 4. </p>
|
||
</strong>
|
||
|
||
|
||
|
||
<div class="figure-caption-contents"><b> The effect of lipin-2 on… </b></div>
|
||
|
||
|
||
|
||
|
||
</div>
|
||
</div>
|
||
|
||
<div class="figure-caption-medium figure-caption-text" aria-hidden="true">
|
||
|
||
|
||
|
||
|
||
|
||
|
||
<strong class="figure-label">
|
||
<p> Figure 4. </p>
|
||
</strong>
|
||
|
||
|
||
|
||
<div class="figure-caption-contents"><b> The effect of lipin-2 on ASC oligomerization and inflammasome assembly. </b> BMDMs from WT…</div>
|
||
|
||
|
||
|
||
|
||
</div>
|
||
|
||
<figcaption id="figure-caption-3" class="figure-caption-full figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="description">
|
||
|
||
|
||
|
||
|
||
|
||
|
||
<strong class="figure-label">
|
||
Figure 4.
|
||
</strong>
|
||
|
||
|
||
|
||
<div class="figure-caption-contents"><b>The effect of lipin-2 on ASC oligomerization and inflammasome assembly.</b> BMDMs from WT and <i>Lpin2<sup>−/−</sup></i> mice (A–D) were treated with 200 ng/ml LPS for 4 h, 2 mM ATP for 40 min, or both, as indicated. (A) Proteins from whole-cell lysates and purified cross-linked ASC oligomers were analyzed by immunoblot using specific antibodies against ASC or β-actin as a loading control. (B and C) Cells were stimulated as indicated and stained with specific antibodies against ASC and DAPI, as mentioned in the Materials and methods. Fluorescence was imaged by confocal microscopy. Bars: (B) 10 µm; (C) 50 µm. Arrowheads denote ASC specks. (D) Fold increase in ASC speck production from 700–1,400 cells is shown. Data from A–D are representative of at least three independent experiments. Data from D are shown as means ± SD *, P < 0.05, Student’s <i>t</i> test.</div>
|
||
|
||
|
||
|
||
|
||
</figcaption>
|
||
|
||
|
||
</figure>
|
||
|
||
<figure class="figure-item "
|
||
itemscope itemtype="http://schema.org/ImageObject"
|
||
itemprop="associatedMedia"
|
||
data-slide-index="4"
|
||
data-label-slug="figure-5">
|
||
<a class="figure-link"
|
||
href="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ba50/5294860/dea28fa9bad3/JEM_20161452_Fig5.jpg"
|
||
itemprop="contentUrl"
|
||
aria-describedby="figure-caption-4"
|
||
role="button"
|
||
data-image-width="781"
|
||
data-image-height="1731"
|
||
data-image-alt="Figure 5."
|
||
data-pmc-id="PMC5294860"
|
||
data-figure-id="fig5">
|
||
<img class="figure-thumb" itemprop="thumbnail"
|
||
id="article-image-4"
|
||
src="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ba50/5294860/6bae930e575d/JEM_20161452_Fig5.gif"
|
||
alt="Figure 5." />
|
||
</a>
|
||
|
||
<meta itemprop="width" itemtype="http://schema.org/ImageObject" content="781">
|
||
<meta itemprop="height" itemtype="http://schema.org/ImageObject" content="1731">
|
||
|
||
|
||
<div class="figure-caption figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="caption" aria-hidden="true">
|
||
|
||
<div class="caption-wrap">
|
||
|
||
|
||
|
||
|
||
|
||
<strong class="figure-label">
|
||
<p> Figure 5. </p>
|
||
</strong>
|
||
|
||
|
||
|
||
<div class="figure-caption-contents"><b> The effect of Lipin-2 on… </b></div>
|
||
|
||
|
||
|
||
|
||
</div>
|
||
</div>
|
||
|
||
<div class="figure-caption-medium figure-caption-text" aria-hidden="true">
|
||
|
||
|
||
|
||
|
||
|
||
|
||
<strong class="figure-label">
|
||
<p> Figure 5. </p>
|
||
</strong>
|
||
|
||
|
||
|
||
<div class="figure-caption-contents"><b> The effect of Lipin-2 on cellular ionic currents generated by ATP. </b> RAW264.7 cells…</div>
|
||
|
||
|
||
|
||
|
||
</div>
|
||
|
||
<figcaption id="figure-caption-4" class="figure-caption-full figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="description">
|
||
|
||
|
||
|
||
|
||
|
||
|
||
<strong class="figure-label">
|
||
Figure 5.
|
||
</strong>
|
||
|
||
|
||
|
||
<div class="figure-caption-contents"><b>The effect of Lipin-2 on cellular ionic currents generated by ATP.</b> RAW264.7 cells treated with control siRNA (siRNACtrl.), siRNA against lipin-2 (siRNA<i>Lpin2</i>; A, left), or BMDMs from WT and <i>Lpin2<sup>−/−</sup></i> mice (A, right) were stimulated with 100 ng/ml LPS for 4 h, 2 mM ATP or 10 µM nigericin for 40 min, 200 µg/ml monosodium urate (MSU) or 150 µg/ml alum for 6 h, or LPS for 4 h, followed by ATP or nigericin for 40 min, MSU, or alum for 6 h, as indicated. IL-1β present in cellular supernatants was analyzed by specific ELISA (A). (B) Representative traces obtained with whole-cell recordings using a 1-s ramp protocol in peritoneal macrophages from WT and <i>Lpin2<sup>−/−</sup></i> animals stimulated with 2 mM ATP in control solution (140 mM NaCl) or in a solution where NaCl has been substituted for 140 mM <i>N</i>-methyl-<sc>d</sc>-glucamine (NMDG), as indicated. (top) Current density versus voltage plots; (bottom) time course of the current density at −120 and +100 mV. (C) Mean current density from 25–39 macrophages analyzed is shown. RAW264.7 cells were also evaluated as in B and mean current density from 18 cells is shown (D). Time course of the current amplitude obtained at −120 mV and +100 mV in a peritoneal macrophage from WT mice upon exposure to 2 mM ATP during the indicated time, to illustrate the calculations of t<sub>on</sub> (time from opening to maximal stimulation) and t<sub>off</sub> (time from maximal opening to closure; E, left). (right) Mean t<sub>on</sub> and t<sub>off</sub> for −120 mV and +100 mV from WT and <i>Lpin2<sup>−/−</sup></i> mice peritoneal macrophages (14–40 cells) stimulated with 2 mM ATP (E). Data from A are shown as mean ± SD, and experiments shown are representative of at least three independent ones made in triplicate. *, P < 0.05, Student’s <i>t</i> test. Dada from C, D, and E, are shown as means ± SEM *, P < 0.05; **, P < 0.01; ***, P < 0.001, one-way ANOVA, followed by Tukey's test.</div>
|
||
|
||
|
||
|
||
|
||
</figcaption>
|
||
|
||
|
||
</figure>
|
||
|
||
<figure class="figure-item "
|
||
itemscope itemtype="http://schema.org/ImageObject"
|
||
itemprop="associatedMedia"
|
||
data-slide-index="5"
|
||
data-label-slug="figure-6">
|
||
<a class="figure-link"
|
||
href="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ba50/5294860/0885bb261bcb/JEM_20161452_Fig6.jpg"
|
||
itemprop="contentUrl"
|
||
aria-describedby="figure-caption-5"
|
||
role="button"
|
||
data-image-width="754"
|
||
data-image-height="599"
|
||
data-image-alt="Figure 6."
|
||
data-pmc-id="PMC5294860"
|
||
data-figure-id="fig6">
|
||
<img class="figure-thumb" itemprop="thumbnail"
|
||
id="article-image-5"
|
||
src="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ba50/5294860/d38928f78576/JEM_20161452_Fig6.gif"
|
||
alt="Figure 6." />
|
||
</a>
|
||
|
||
<meta itemprop="width" itemtype="http://schema.org/ImageObject" content="754">
|
||
<meta itemprop="height" itemtype="http://schema.org/ImageObject" content="599">
|
||
|
||
|
||
<div class="figure-caption figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="caption" aria-hidden="true">
|
||
|
||
<div class="caption-wrap">
|
||
|
||
|
||
|
||
|
||
|
||
<strong class="figure-label">
|
||
<p> Figure 6. </p>
|
||
</strong>
|
||
|
||
|
||
|
||
<div class="figure-caption-contents"><b> Effect of lipin-2 on ATP-induced… </b></div>
|
||
|
||
|
||
|
||
|
||
</div>
|
||
</div>
|
||
|
||
<div class="figure-caption-medium figure-caption-text" aria-hidden="true">
|
||
|
||
|
||
|
||
|
||
|
||
|
||
<strong class="figure-label">
|
||
<p> Figure 6. </p>
|
||
</strong>
|
||
|
||
|
||
|
||
<div class="figure-caption-contents"><b> Effect of lipin-2 on ATP-induced changes in K <sup> + </sup> cellular levels and cell… </b></div>
|
||
|
||
|
||
|
||
|
||
</div>
|
||
|
||
<figcaption id="figure-caption-5" class="figure-caption-full figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="description">
|
||
|
||
|
||
|
||
|
||
|
||
|
||
<strong class="figure-label">
|
||
Figure 6.
|
||
</strong>
|
||
|
||
|
||
|
||
<div class="figure-caption-contents"><b>Effect of lipin-2 on ATP-induced changes in K<sup>+</sup> cellular levels and cell permeability.</b> (A) Control (siRNACtrl.) and lipin-2–silenced (siRNA<i>Lpin2</i>) RAW264.7 cells were stimulated with 2 mM ATP for the indicated periods of time, and intracellular concentration of K<sup>+</sup> was analyzed using an inductively coupled plasma/optical emission spectrometer. (B) RAW264.7 cells were stimulated with 200 ng/ml LPS for 4 h, and then 2 mM ATP for 40 min, as indicated, in the presence of 0, 5, or 45 mM K<sup>+</sup>. IL-1β present in cellular supernatants was analyzed by specific ELISAS. (C) BMDMs from WT and <i>Lpin2<sup>−/−</sup></i> mice were pretreated with 20 µM ethidium bromide (BrEt) for 5 min, and then stimulated with 2 mM ATP. Ethidium cellular uptake was monitored by confocal microscopy. (top) Cell fluorescence; (bottom) fluorescence fold increase over time. Bar, 25 µm. (D) Control (siRNACtrl.) and lipin-2–silenced (siRNA<i>Lpin2</i>) RAW264.7 cells were stimulated with 2 mM ATP for 50 min in the presence of Annexin-V-Cy3, and fluorescence analyzed in a confocal microscope (top). (bottom) Fluorescence fold increase over unstimulated cells. Bar, 50 µm. Data from A and B are shown as mean ± SD, and experiments shown are representative of at least three independent experiments made in triplicates. Data from C (>100 cells) and D (>300 cells) are shown as means ± SEM, and experiments shown are representative of at least three independent ones. *, P < 0.05; **, P < 0.01; ***, P < 0.001, by Student’s <i>t</i> test.</div>
|
||
|
||
|
||
|
||
|
||
</figcaption>
|
||
|
||
|
||
</figure>
|
||
|
||
<figure class="figure-item tail"
|
||
itemscope itemtype="http://schema.org/ImageObject"
|
||
itemprop="associatedMedia"
|
||
data-slide-index="6"
|
||
data-label-slug="figure-7">
|
||
<a class="figure-link"
|
||
href="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ba50/5294860/7dc68b6c0015/JEM_20161452_Fig7.jpg"
|
||
itemprop="contentUrl"
|
||
aria-describedby="figure-caption-6"
|
||
role="button"
|
||
data-image-width="782"
|
||
data-image-height="871"
|
||
data-image-alt="Figure 7."
|
||
data-pmc-id="PMC5294860"
|
||
data-figure-id="fig7">
|
||
<img class="figure-thumb" itemprop="thumbnail"
|
||
id="article-image-6"
|
||
src="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ba50/5294860/a9740cf56c48/JEM_20161452_Fig7.gif"
|
||
alt="Figure 7." />
|
||
</a>
|
||
|
||
<meta itemprop="width" itemtype="http://schema.org/ImageObject" content="782">
|
||
<meta itemprop="height" itemtype="http://schema.org/ImageObject" content="871">
|
||
|
||
|
||
<div class="figure-caption figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="caption" aria-hidden="true">
|
||
|
||
<div class="caption-wrap">
|
||
|
||
|
||
|
||
|
||
|
||
<strong class="figure-label">
|
||
<p> Figure 7. </p>
|
||
</strong>
|
||
|
||
|
||
|
||
<div class="figure-caption-contents"><b> Impact of lipin-2 on cellular… </b></div>
|
||
|
||
|
||
|
||
|
||
</div>
|
||
</div>
|
||
|
||
<div class="figure-caption-medium figure-caption-text" aria-hidden="true">
|
||
|
||
|
||
|
||
|
||
|
||
|
||
<strong class="figure-label">
|
||
<p> Figure 7. </p>
|
||
</strong>
|
||
|
||
|
||
|
||
<div class="figure-caption-contents"><b> Impact of lipin-2 on cellular cholesterol levels, and restoration of P2X <sub> 7 </sub> R… </b></div>
|
||
|
||
|
||
|
||
|
||
</div>
|
||
|
||
<figcaption id="figure-caption-6" class="figure-caption-full figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="description">
|
||
|
||
|
||
|
||
|
||
|
||
|
||
<strong class="figure-label">
|
||
Figure 7.
|
||
</strong>
|
||
|
||
|
||
|
||
<div class="figure-caption-contents"><b>Impact of lipin-2 on cellular cholesterol levels, and restoration of P2X<sub>7</sub>R functionality by cholesterol in the absence of lipin-2.</b> (A) BMDMs from WT and <i>Lpin2<sup>−/−</sup></i> mice (left), or control (siRNACtrl.) and lipin-2–silenced (siRNA<i>Lpin2</i>) RAW264.7 cells (right) were treated with 200 ng/ml LPS for 4 h, 2 mM ATP for 40 min, or both, as indicated. (B) Total cellular cholesterol levels were analyzed as described in Materials and methods. RAW264.7 cells preincubated with 100 µg/ml cholesterol (cholesterol/MCD) for 30 min were treated as in A, and total cellular cholesterol levels were measured. (C) Example traces of whole-cell ionic currents analyzed by patch-clamp in peritoneal macrophages from WT and <i>Lpin2<sup>−/−</sup></i> animals stimulated with 2 mM ATP in the absence or presence of 100 µg/ml cholesterol (cholesterol/MCD) as indicated. (top) Current density against voltage; (bottom) current densities at −120 and +100 mV. (D) Mean current density from 15–18 cells analyzed. (E) The mean t<sub>on</sub> and t<sub>off</sub> at −120 and +100 mV from WT and <i>Lpin2<sup>−/−</sup></i> mice peritoneal macrophages stimulated with 2 mM ATP in the presence or absence of 100 µg/ml cholesterol (cholesterol/MCD). Data from A and B are shown as mean ± SD, and experiments shown are representative of at least three independent experiments made in triplicate. Data from D and E are shown as means ± SEM *, P < 0.05; **, P < 0.01; ***, P < 0.001, Student’s <i>t</i> test (A and B) or by one-way ANOVA followed by Tukey's test (D and E).</div>
|
||
|
||
|
||
|
||
|
||
</figcaption>
|
||
|
||
|
||
</figure>
|
||
|
||
<figure class="figure-item tail"
|
||
itemscope itemtype="http://schema.org/ImageObject"
|
||
itemprop="associatedMedia"
|
||
data-slide-index="7"
|
||
data-label-slug="figure-8">
|
||
<a class="figure-link"
|
||
href="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ba50/5294860/c222e5546415/JEM_20161452_Fig8.jpg"
|
||
itemprop="contentUrl"
|
||
aria-describedby="figure-caption-7"
|
||
role="button"
|
||
data-image-width="760"
|
||
data-image-height="519"
|
||
data-image-alt="Figure 8."
|
||
data-pmc-id="PMC5294860"
|
||
data-figure-id="fig8">
|
||
<img class="figure-thumb" itemprop="thumbnail"
|
||
id="article-image-7"
|
||
src="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ba50/5294860/3636b31edc67/JEM_20161452_Fig8.gif"
|
||
alt="Figure 8." />
|
||
</a>
|
||
|
||
<meta itemprop="width" itemtype="http://schema.org/ImageObject" content="760">
|
||
<meta itemprop="height" itemtype="http://schema.org/ImageObject" content="519">
|
||
|
||
|
||
<div class="figure-caption figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="caption" aria-hidden="true">
|
||
|
||
<div class="caption-wrap">
|
||
|
||
|
||
|
||
|
||
|
||
<strong class="figure-label">
|
||
<p> Figure 8. </p>
|
||
</strong>
|
||
|
||
|
||
|
||
<div class="figure-caption-contents"><b> Cholesterol effect on ASC oligomerization,… </b></div>
|
||
|
||
|
||
|
||
|
||
</div>
|
||
</div>
|
||
|
||
<div class="figure-caption-medium figure-caption-text" aria-hidden="true">
|
||
|
||
|
||
|
||
|
||
|
||
|
||
<strong class="figure-label">
|
||
<p> Figure 8. </p>
|
||
</strong>
|
||
|
||
|
||
|
||
<div class="figure-caption-contents"><b> Cholesterol effect on ASC oligomerization, capase-1 activation, and IL-1β production in the absence… </b></div>
|
||
|
||
|
||
|
||
|
||
</div>
|
||
|
||
<figcaption id="figure-caption-7" class="figure-caption-full figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="description">
|
||
|
||
|
||
|
||
|
||
|
||
|
||
<strong class="figure-label">
|
||
Figure 8.
|
||
</strong>
|
||
|
||
|
||
|
||
<div class="figure-caption-contents"><b>Cholesterol effect on ASC oligomerization, capase-1 activation, and IL-1β production in the absence of lipin-2.</b> (A) BMDMs from WT and <i>Lpin2<sup>−/−</sup></i> mice were treated with 200 ng/ml LPS for 4 h, 2 mM ATP, or both, as indicated. Some samples were preincubated with 100 µg/ml cholesterol 30 min before ATP treatment, as indicated. Proteins from whole-cell lysates and purified cross-linked ASC oligomers were analyzed by immunoblot using specific antibodies against ASC or β-actin, as a loading control. (B) Control (siRNACtrl.) or lipin-2–silenced (siRNA<i>Lpin2</i>) RAW264.7 cells were treated with 200 ng/ml LPS for 4 h and then 2 mM ATP for 40 min, as indicated. Some samples were preincubated with 100 µg/ml cholesterol (cholesterol/MCD) 30 min before ATP treatment, as indicated. Intracellular active caspase was analyzed by flow cytometry as specified in Materials and methods. Median fluorescence intensities (MFI) are indicated. (bottom) Percentage of cells with active caspase-1. (C) RAW264.7 cells were treated as in B and presence of IL-1β in cellular supernatants was quantified by specific ELISA in triplicate samples. Data are shown as means ± SD, and experiments shown are representative of at least three independent experiments. *, P < 0.05; **, P < 0.01, by Student’s <i>t</i> test.</div>
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<p> Figure 9. </p>
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<div class="figure-caption-contents"><b> The effect of lipin-2 in… </b></div>
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<p> Figure 9. </p>
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<div class="figure-caption-contents"><b> The effect of lipin-2 in the response to LPS in mice. </b> (A) WT…</div>
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Figure 9.
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<div class="figure-caption-contents"><b>The effect of lipin-2 in the response to LPS in mice.</b> (A) WT and <i>Lpin2<sup>−/−</sup></i> mice (<i>n</i> = 5) were intraperitoneally injected with 10 mg/kg of LPS or PBS. 3 h later, indicated cytokines were quantified in serum using specific ELISA. Expression levels of the indicated genes were analyzed in liver (B) or spleen (C) by RT-qPCR. (D) Proposed scheme depicting lipin-2 effects on priming and activation of the inflammasome. Data are shown as means ± SD, and experiments shown are representative of at least two independent ones analyzed in triplicates. *, P < 0.05; **, P < 0.01; ***, P < 0.001, by Student’s <i>t</i> test.</div>
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Al-Mosawi, Z.S., Al-Saad K.K., Ijadi-Maghsoodi R., El-Shanti H.I., and Ferguson P.J.. 2007. A splice site mutation confirms the role of LPIN2 in Majeed syndrome. Arthritis Rheum. 56:960–964. 10.1002/art.22431
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NLR Family, Pyrin Domain-Containing 3 Protein / physiology*
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Receptors, Purinergic P2X7
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