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</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="lR1CjY2NbnvMyMM5Mv2eCHUKRAMZwVd9CELHWOv9LYigsdSVpOFg6PqopEubpwLt">
<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="[^&quot;&amp;=&lt;&gt;\/]*" title="The following characters are not allowed in the Name field: &quot;&amp;=&lt;&gt;/"
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="lR1CjY2NbnvMyMM5Mv2eCHUKRAMZwVd9CELHWOv9LYigsdSVpOFg6PqopEubpwLt">
<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="lR1CjY2NbnvMyMM5Mv2eCHUKRAMZwVd9CELHWOv9LYigsdSVpOFg6PqopEubpwLt">
<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="[^&quot;&amp;=&lt;&gt;\/]*" title="The following characters are not allowed in the Name field: &quot;&amp;=&lt;&gt;/">
</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="lR1CjY2NbnvMyMM5Mv2eCHUKRAMZwVd9CELHWOv9LYigsdSVpOFg6PqopEubpwLt">
<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="lR1CjY2NbnvMyMM5Mv2eCHUKRAMZwVd9CELHWOv9LYigsdSVpOFg6PqopEubpwLt">
<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="[^&quot;&amp;=&lt;&gt;\/]*" title="The following characters are not allowed in the Name field: &quot;&amp;=&lt;&gt;/">
</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="21282379">
<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.20102049"
target="_blank"
rel="noopener"
ref="linksrc=fulltextorjournal_fulltext&amp;is_pmc=False&amp;PrId=7898&amp;itool=Abstract-def&amp;log$=linkouticon&amp;uid=21282379&amp;db=pubmed&amp;nlmid=2985109R"
title="See full text options at Silverchair Information Systems"
data-ga-category="full_text"
data-ga-action="Silverchair Information Systems"
data-ga-label="21282379"
><img src="https://cdn.ncbi.nlm.nih.gov/corehtml/query/egifs/https:--rupress.org-images-pubmed-jem_free.png" alt="Silverchair Information Systems full text link"><span class="text">
Silverchair Information Systems
</span></a><a class="link-item
pmc
"
href="https://pmc.ncbi.nlm.nih.gov/articles/pmid/21282379/"
target="_blank"
rel="noopener"
ref="linksrc=fulltextorjournal_fulltext&amp;is_pmc=True&amp;PrId=3494&amp;itool=Abstract-def&amp;log$=linkouticon&amp;uid=21282379&amp;db=pubmed&amp;nlmid=2985109R"
title="Free full text at PubMed Central"
data-ga-category="full_text"
data-ga-action="PMC"
data-ga-label="21282379"
><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="/21282379/citations/"
data-citation-style="nlm"
data-pubmed-format-link="/21282379/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="21282379" 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%2F21282379%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="lR1CjY2NbnvMyMM5Mv2eCHUKRAMZwVd9CELHWOv9LYigsdSVpOFg6PqopEubpwLt"><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"
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Abstract
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<p>
Reactive oxygen species (ROS) have an established role in inflammation and host defense, as they kill intracellular bacteria and have been shown to activate the NLRP3 inflammasome. Here, we find that ROS generated by mitochondrial respiration are important for normal lipopolysaccharide (LPS)-driven production of several proinflammatory cytokines and for the enhanced responsiveness to LPS seen in cells from patients with tumor necrosis factor receptor-associated periodic syndrome (TRAPS), an autoinflammatory disorder caused by missense mutations in the type 1 TNF receptor (TNFR1). We find elevated baseline ROS in both mouse embryonic fibroblasts and human immune cells harboring TRAPS-associated TNFR1 mutations. A variety of antioxidants dampen LPS-induced MAPK phosphorylation and inflammatory cytokine production. However, gp91(phox) and p22(phox) reduced nicotinamide adenine dinucleotide phosphate (NADPH) oxidase subunits are dispensable for inflammatory cytokine production, indicating that NADPH oxidases are not the source of proinflammatory ROS. TNFR1 mutant cells exhibit altered mitochondrial function with enhanced oxidative capacity and mitochondrial ROS generation, and pharmacological blockade of mitochondrial ROS efficiently reduces inflammatory cytokine production after LPS stimulation in cells from TRAPS patients and healthy controls. These findings suggest that mitochondrial ROS may be a novel therapeutic target for TRAPS and other inflammatory diseases.
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data-image-width="411"
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data-image-alt="Figure 1."
data-pmc-id="PMC3058571"
data-figure-id="fig1">
<img class="figure-thumb" itemprop="thumbnail"
id="article-image-0"
src="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1c08/3058571/4cb2b49c2a7b/JEM_20102049_RGB_Fig1.gif"
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<strong class="figure-label">
<p> Figure 1. </p>
</strong>
<div class="figure-caption-contents"><b> Increased ROS in TNFR1 mutant… </b></div>
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<div class="figure-caption-medium figure-caption-text" aria-hidden="true">
<strong class="figure-label">
<p> Figure 1. </p>
</strong>
<div class="figure-caption-contents"><b> Increased ROS in TNFR1 mutant cells and peripheral blood cells from TRAPS patients.… </b></div>
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<figcaption id="figure-caption-0" class="figure-caption-full figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="description">
<strong class="figure-label">
Figure 1.
</strong>
<div class="figure-caption-contents"><b>Increased ROS in TNFR1 mutant cells and peripheral blood cells from TRAPS patients.</b> (A) WT, C33Y TNFR1 heterozygous, and T50M TNFR1 heterozygous MEFs were left untreated (UT) or were treated with PMA or LPS for 1 h. Cells were incubated with DHR and ROS levels were determined by flow cytometry. Bar graph shows results of four independent experiments and histograms show one representative experiment. n.s., not significant. Error bars represent the mean ± SEM. (B) Monocytes and neutrophils from healthy donors (<i>n</i> = 2731), TRAPS patients with structural mutations (TRAPS structural; <i>n</i> = 1113), TRAPS patients with R92Q polymorphisms (TRAPS poly; <i>n</i> = 6), or patients with familial Mediterranean fever (FMF; <i>n</i> = 7) were treated for 1 h with or without PMA. Cells were incubated with DHR, and ROS levels were determined by flow cytometry. Data are from 10 experiments; each symbol represents a unique patient. (C) PBMCs and neutrophils from healthy donors (<i>n</i> = 2533), TRAPS patients with structural mutation (TRAPS structural; <i>n</i> = 1316) and TRAPS patients with R92Q or P46L polymorphisms (TRAPS poly; <i>n</i> = 4) were analyzed for superoxide production with a kinetic chemiluminescent assay in the presence or absence of PMA. Data are from 11 experiments; each symbol represents a unique patient. RLU, relative luminescence units. Data were normalized to the mean WT or healthy donor result for each experiment. *, P ≤ 0.05; **, P &lt; 0.01.</div>
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<img class="figure-thumb" itemprop="thumbnail"
id="article-image-1"
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<strong class="figure-label">
<p> Figure 2. </p>
</strong>
<div class="figure-caption-contents"><b> Role of ROS in LPS-induced… </b></div>
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<div class="figure-caption-medium figure-caption-text" aria-hidden="true">
<strong class="figure-label">
<p> Figure 2. </p>
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<div class="figure-caption-contents"><b> Role of ROS in LPS-induced MAPK activation and cytokine production in normal and… </b></div>
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<figcaption id="figure-caption-1" class="figure-caption-full figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="description">
<strong class="figure-label">
Figure 2.
</strong>
<div class="figure-caption-contents"><b>Role of ROS in LPS-induced MAPK activation and cytokine production in normal and TNFR1 mutant cells.</b> (A) WT and C33Y TNFR1 heterozygous MEFs were treated with or without NAC for 30 min, and further treated with LPS for the indicated time, after which JNK, p38, and ERK phosphorylation were measured by Western blot. Numbers shown are the density of each phosphoprotein relative to nonphosphorylated protein, normalized to the WT untreated sample. Actin is included as a loading control. Data are representative of three independent experiments. (B) WT, C33Y TNFR1 heterozygous, and T50M TNFR1 heterozygous MEFs were treated with LPS in the presence of the indicated antioxidants. IL-6 and IFN-β were measured in supernatants. Levels produced by WT MEFs after LPS stimulation ranged from 11.5104.5 pg/ml (IL-6) and 13.523.0 pg/ml (IFN-β; <i>n</i> = 4 independent experiments). Data were normalized to the average WT result for each experiment. (C) IL-6 mRNA was measured by qRT-PCR in MEFs of the indicated genotype treated for 6 h with LPS and the indicated antioxidants, as in B. Data are shown as fold induction to LPS (as compared with untreated), and are representative of three experiments. (D) PBMCs from healthy donors (<i>n</i> = 56) and TRAPS patients with structural mutation (<i>n</i> = 47) were incubated with LPS and the indicated antioxidants, as in B. IL-6, TNF, CXCL8/IL-8, IL-10, IFN-β, and CCL5/RANTES were measured in supernatants. Levels produced by cells from healthy donors after LPS stimulation ranged from 2.369.98 ng/ml (IL-6), 0.212.04 ng/ml (TNF), 5.777.7 ng/ml (CXCL8/IL-8), 0.0540.334 ng/ml (IL-10), 0.0100.058 ng/ml (IFN-β), and 0.1380.979 ng/ml (CCL5/RANTES; <i>n</i> = 35 independent experiments). Error bars represent the mean ± SEM. Data were normalized to the average healthy donor result for each experiment. *, P ≤ 0.05; **, P &lt; 0.01.</div>
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data-pmc-id="PMC3058571"
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<img class="figure-thumb" itemprop="thumbnail"
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alt="Figure 3." />
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<strong class="figure-label">
<p> Figure 3. </p>
</strong>
<div class="figure-caption-contents"><b> NOX-derived ROS and the inflammasome… </b></div>
</div>
</div>
<div class="figure-caption-medium figure-caption-text" aria-hidden="true">
<strong class="figure-label">
<p> Figure 3. </p>
</strong>
<div class="figure-caption-contents"><b> NOX-derived ROS and the inflammasome are not required for LPS-induced cytokine production. </b> (A)…</div>
</div>
<figcaption id="figure-caption-2" class="figure-caption-full figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="description">
<strong class="figure-label">
Figure 3.
</strong>
<div class="figure-caption-contents"><b>NOX-derived ROS and the inflammasome are not required for LPS-induced cytokine production.</b> (A) Peritoneal macrophages from mice of the indicated genotypes were stimulated with LPS for 6 h, and ATP for the final 15 min, before measuring IL-6, TNF, and IL-1β in the supernatants. Levels produced by WT mice after LPS stimulation ranged from 1.521.61 ng/ml (IL-6), 0.230.97 ng/ml (TNF), and 0.210.46 ng/ml (IL-1β; <i>n</i> = 2 independent experiments, 1 mouse in each group). (B) Peritoneal macrophages from mice of the indicated genotypes were incubated with LPS in the presence or absence of DPI for 6 h, and ATP for the final 15 min, before measuring IL-6, TNF, and IL-1β in the supernatants. Levels produced by WT mice after LPS stimulation ranged from 1.524.49 ng/ml (IL-6), 0.140.19 ng/ml (TNF), and 3.104.07 ng/ml (IL-1β; <i>n</i> = 3 independent experiments, one mouse in each group). (C) Peritoneal macrophages from mice of the indicated genotypes were stimulated with LPS for 6 h, and ATP for the final 15 min, before measuring IL-6, TNF, and IL-1β in the supernatants. Levels produced by WT mice after LPS stimulation ranged from 1.682.70 ng/ml (IL-6), 0.390.58 ng/ml (TNF), and 0.180.80 ng/ml (IL-1β; <i>n</i> = 3 independent experiments, one mouse in each group). nd, not detectable. Error bars represent the mean ± SEM. Data were normalized to the average WT LPS result for each experiment. *, P ≤ 0.05; **, P &lt; 0.01.</div>
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data-image-alt="Figure 4."
data-pmc-id="PMC3058571"
data-figure-id="fig4">
<img class="figure-thumb" itemprop="thumbnail"
id="article-image-3"
src="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1c08/3058571/90b9982baf32/JEM_20102049_GS_Fig4.gif"
alt="Figure 4." />
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<strong class="figure-label">
<p> Figure 4. </p>
</strong>
<div class="figure-caption-contents"><b> Inhibition of mitochondrial oxygen consumption… </b></div>
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<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> Inhibition of mitochondrial oxygen consumption by DPI. </b> (A) The effect of antioxidants on…</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>Inhibition of mitochondrial oxygen consumption by DPI.</b> (A) The effect of antioxidants on oxygen consumption rate was measured in WT MEFs with a Seahorse Bioscience XF Analyzer. The arrow indicates time of addition of the antioxidants. A representative plot (<i>n</i> = 3) is shown. (B) WT MEFs were incubated with the indicated antioxidants for 1 h or were left untreated and then stained with MitoSOX Red, and ROS levels were determined by flow cytometry. Data were normalized to the untreated result (<i>n</i> = 3 independent experiments). (C) WT MEFs were treated with LPS and the indicated concentrations of DPI, after which IL-6 was measured in supernatants. Levels of IL-6 were normalized to the LPS alone result, which ranged from 0.2840.321 ng/ml (<i>n</i> = 3 independent experiments). (D) WT MEFs were treated with PMA and the indicated concentrations of DPI for 1 h. Cells were incubated with DHR, and ROS levels were determined by flow cytometry. Data were normalized to the increase in fluorescence (as compared with untreated) of the PMA alone sample for each experiment (<i>n</i> = 3 independent experiments). (E) WT, C33Y TNFR1 heterozygous, and T50M TNFR1 heterozygous MEFs were treated with LPS in the presence or absence of rotenone, after which IL-6 was measured in supernatants. IL-6 levels were normalized to the WT LPS alone result, which ranged from 11.5104.5 pg/ml (<i>n</i> = 4 independent experiments). (F) PBMCs from healthy donors (<i>n</i> = 6) and TRAPS patients with structural mutations (<i>n</i> = 6) were incubated with LPS in the presence or absence of rotenone, after which IL-6 and TNF were measured in the supernatants. IL-6 and TNF levels were normalized to the healthy donor LPS alone result, which ranged from 1.312.28 ng/ml (IL-6) and 0.2740.661 ng/ml (TNF; <i>n</i> = 6 independent experiments). Error bars represent the mean ± SEM.</div>
</figcaption>
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data-image-width="448"
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data-image-alt="Figure 5."
data-pmc-id="PMC3058571"
data-figure-id="fig5">
<img class="figure-thumb" itemprop="thumbnail"
id="article-image-4"
src="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1c08/3058571/3db6061ead2e/JEM_20102049_GS_Fig5.gif"
alt="Figure 5." />
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<strong class="figure-label">
<p> Figure 5. </p>
</strong>
<div class="figure-caption-contents"><b> Increased ATP levels and oxidative… </b></div>
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</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> Increased ATP levels and oxidative capacity in cells with TRAPS-associated TNFR1 mutations. </b> (A)…</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>Increased ATP levels and oxidative capacity in cells with TRAPS-associated TNFR1 mutations.</b> (A) ATP level was measured in WT and TNFR1 mutant MEFs. Cell numbers were determined with a CyQuant assay, and data were normalized to the WT result (<i>n</i> = 5 independent experiments). RLU, relative luminescence units. (B) ATP level was measured in PBMCs from healthy donors (<i>n</i> = 8) and TRAPS patients with structural mutations (<i>n</i> = 6). Cell numbers were determined with a CyQuant assay, and data were normalized to the healthy donor result (<i>n</i> = 4 independent experiments). RLU, relative luminescence units. (C) Oxygen consumption was measured in WT, C33Y TNFR1 heterozygous, and T50M TNFR1 heterozygous MEFs using the Seahorse Bioscience XF Analyzer. Dinitrophenol was added as indicated by the arrow. A representative plot (<i>n</i> = 3) is shown. (D) Basal and maximum oxygen consumption were determined in MEFs of the indicated genotypes using the Seahorse Bioscience XF Analyzer (<i>n</i> = 3 independent experiments). (E) TMRM staining of WT, C33Y TNFR1 heterozygous, and T50M TNFR1 heterozygous MEFs was determined by flow cytometry. A representative histogram (<i>n</i> = 3) is shown. (F) Extracellular acidification was measured in WT, C33Y TNFR1 heterozygous, and T50M TNFR1 heterozygous MEFs using the Seahorse Bioscience XF Analyzer (<i>n</i> = 3 independent experiments). (G) Mitochondrial copy number was measured by qRT-PCR in MEFs of the indicated genotypes. Cytb mitochondrial DNA was compared with 18S genomic DNA (<i>n</i> = 2 independent experiments). (H) Oxygen consumption was measured in PBMCs from healthy donors (<i>n</i> = 10) and patients with TRAPS structural mutations (<i>n</i> = 6) using the Seahorse Bioscience XF Analyzer. Dinitrophenol was added as indicated by the arrow. A representative plot with two healthy donors and two TRAPS patients with structural mutations is shown (<i>n</i> = 5 independent experiments). (I) Basal and maximum oxygen consumption were determined from healthy donors (<i>n</i> = 10) and TRAPS patients with structural mutations (TRAPS structural; <i>n</i> = 6) using the Seahorse Bioscience XF Analyzer. Error bars represent the mean ± SEM. Data were normalized to the average healthy donor result (<i>n</i> = 5 independent experiments). *, P ≤ 0.05; **, P &lt; 0.01.</div>
</figcaption>
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data-image-alt="Figure 6."
data-pmc-id="PMC3058571"
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<strong class="figure-label">
<p> Figure 6. </p>
</strong>
<div class="figure-caption-contents"><b> Mitochondrial superoxide production is increased… </b></div>
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</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> Mitochondrial superoxide production is increased in TRAPS. </b> (A) WT, C33Y TNFR1 heterozygous, and…</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>Mitochondrial superoxide production is increased in TRAPS.</b> (A) WT, C33Y TNFR1 heterozygous, and T50M TNFR1 heterozygous MEFs were incubated with MitoSOX Red for 1 h and ROS levels were determined by flow cytometry. Histogram shows representative staining and bar graph shows results of three independent experiments. Data were normalized to the average WT result for each experiment. (B) Monocytes from healthy donors (<i>n</i> = 15), TRAPS patients with structural mutations (TRAPS structural; <i>n</i> = 9), and TRAPS patients with TNFR1 polymorphisms (TRAPS poly; <i>n</i> = 5) were incubated with MitoSOX Red for 1 h, and ROS levels were determined by flow cytometry. Histogram shows representative staining and bar graph shows results of six independent experiments; each symbol represents a unique patient. Data were normalized to the average healthy donor result for each experiment. (C) SOD2 mRNA was measured by qRT-PCR in MEFs of the indicated genotype (<i>n</i> = 3 independent experiments). Data represent gene expression relative to β2-microglobulin. (D) SOD2 and thioredoxin mRNA were measured by qRT-PCR in PBMCs from healthy donors (<i>n</i> = 67) and TRAPS patients with structural mutations (<i>n</i> = 14). Data represent gene expression relative to β2-microglobulin. Error bars represent the mean ± SEM. *, P ≤ 0.05; **, P &lt; 0.01.</div>
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alt="Figure 7." />
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<div class="figure-caption-contents"><b> Inhibition of mitochondrial ROS can… </b></div>
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<div class="figure-caption-contents"><b> Inhibition of mitochondrial ROS can reduce normal cytokine production and reverse hyperinflammatory responses… </b></div>
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Figure 7.
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<div class="figure-caption-contents"><b>Inhibition of mitochondrial ROS can reduce normal cytokine production and reverse hyperinflammatory responses in TRAPS.</b> (A) WT, C33Y TNFR1 heterozygous, and T50M TNFR1 heterozygous MEFs were treated with LPS in the presence of the mitochondrial ROS scavenger MitoQ and its control, dTPP. IL-6 and IFN-β were measured in supernatants. Levels produced by WT MEFs after LPS stimulation ranged from 11.5104.5 pg/ml (IL-6) and 13.523.0 pg/ml (IFN-β; <i>n</i> = 3 independent experiments). (B) IL-6 mRNA was measured by qRT-PCR in MEFs of the indicated genotype treated for 6 h with LPS and the indicated reagents as in A. Data are shown as fold induction to LPS (as compared with untreated), and are representative of three independent experiments. (C) PBMCs from healthy donors (<i>n</i> = 58) and TRAPS patients with structural mutations (TRAPS structural; <i>n</i> = 4) were incubated with LPS and the indicated reagents as in A. IL-6, TNF, CXCL8/IL-8, IL-10, IFN-β, and CCL5/RANTES were measured in supernatants. Levels produced by PBMCs from healthy donors after LPS stimulation ranged from 0.938.67 ng/ml (IL-6), 1.174.12 ng/ml (TNF), 5.777.7 ng/ml (CXCL8/IL-8), 0.0540.334 ng/ml (IL-10), 0.0100.058 ng/ml (IFN-β), and 0.1380.979 ng/ml (CCL5/RANTES; <i>n</i> = 34 independent experiments). Data were normalized to the average WT or healthy donor LPS result for each experiment. *, P ≤ 0.05; **, P &lt; 0.01. (D) Schematic model of how mitochondrial ROS support LPS hyperresponsiveness in TRAPS.</div>
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