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<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="W58aGdkXdPIjEJDUSYuGd40kPtZWPNFRdSSfj3NjNqvNyaJKvh7IHcwYnxH8Iodb">
<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="W58aGdkXdPIjEJDUSYuGd40kPtZWPNFRdSSfj3NjNqvNyaJKvh7IHcwYnxH8Iodb">
<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="W58aGdkXdPIjEJDUSYuGd40kPtZWPNFRdSSfj3NjNqvNyaJKvh7IHcwYnxH8Iodb">
<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="W58aGdkXdPIjEJDUSYuGd40kPtZWPNFRdSSfj3NjNqvNyaJKvh7IHcwYnxH8Iodb">
<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="W58aGdkXdPIjEJDUSYuGd40kPtZWPNFRdSSfj3NjNqvNyaJKvh7IHcwYnxH8Iodb">
<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="19017994">
<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://journals.aai.org/jimmunol/article-lookup/doi/10.4049/jimmunol.181.11.8018"
target="_blank"
rel="noopener"
ref="linksrc=fulltextorjournal_fulltext&amp;is_pmc=False&amp;PrId=7898&amp;itool=Abstract-def&amp;log$=linkouticon&amp;uid=19017994&amp;db=pubmed&amp;nlmid=2985117R"
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Silverchair Information Systems
</span></a><a class="link-item
pmc
"
href="https://pmc.ncbi.nlm.nih.gov/articles/pmid/19017994/"
target="_blank"
rel="noopener"
ref="linksrc=fulltextorjournal_fulltext&amp;is_pmc=True&amp;PrId=3494&amp;itool=Abstract-def&amp;log$=linkouticon&amp;uid=19017994&amp;db=pubmed&amp;nlmid=2985117R"
title="Free full text at PubMed Central"
data-ga-category="full_text"
data-ga-action="PMC"
data-ga-label="19017994"
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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="/19017994/citations/"
data-citation-style="nlm"
data-pubmed-format-link="/19017994/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="19017994" 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%2F19017994%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="W58aGdkXdPIjEJDUSYuGd40kPtZWPNFRdSSfj3NjNqvNyaJKvh7IHcwYnxH8Iodb"><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:
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SOCS1 regulates the IFN but not NFkappaB pathway in TLR-stimulated human monocytes and macrophages
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<p>
SOCS1 can regulate TLR-mediated signal transduction, yet mechanistic studies in murine macrophages have been confusing and contradictory. This study has used an adenoviral transfection system to determine the role of SOCS1 in the regulation of TNF-alpha production by activated human monocytes. Monocytes were infected with AdV-SOCS1 or with an empty vector control, AdV-GFP, for 24 h before activation with the TLR4 ligand, LPS. SOCS1 did not regulate TNF-alpha mRNA or protein production within the first two hours of TLR4 activation. However, SOCS1 suppressed the sustained production of TNF-alpha by primary human monocytes and synovial fluid macrophages ex vivo. In addition, SOCS1 regulated the production of IL-6, but not IL-10, by monocytes. Analysis of the early signaling pathway downstream of TLR4 demonstrated that SOCS1 had no regulatory effect on the activation or on the DNA binding capacity of NFkappaB. The late effects of LPS are mediated in part through the MyD88-independent pathway activating IRF3 and initiating the production of IFN-beta. In response to adenoviral infection and before LPS exposure, monocytes expressed enhanced levels of IFN-beta and Myxovirus A mRNA, an anti-viral molecule characterizing IFN-beta activity. These two genes were reduced in AdV-SOCS1-infected cells. Further, SOCS1 regulated IFN-dependent pathways in LPS-activated cells as evidenced by reduced IFN-beta production and STAT1 phosphorylation. Using AdV-infection to dissect SOCS1 control of IFN-dependent pathways, this study suggests that SOCS1-regulation of the IFN-dependent component of the LPS-induced TLR4 signaling pathway may contribute to the down-regulation of inflammatory cytokine production by AdV-SOCS1-infected human monocytes.
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<p> FIGURE 1. SOCS1 expression by AdV-SOCS1-infected human… </p>
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<p> FIGURE 1. SOCS1 expression by AdV-SOCS1-infected human monocytes </p>
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<div class="figure-caption-contents"><i> A. </i> LPS induced SOCS1 mRNA levels over…</div>
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FIGURE 1. SOCS1 expression by AdV-SOCS1-infected human monocytes
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<div class="figure-caption-contents"><i>A.</i> LPS induced SOCS1 mRNA levels over 2 h in monocytes from each of 4 individual donors (mean ± SEM, n=3 replicates/donor). *p&lt;0.05 compared to untreated (time 0), 0.5 and 1 h following LPS exposure. SOCS1 mRNA was normalized to the housekeeping gene UBE2D2. <i>B.</i> Infection efficiency (% GFP positive cells, determined using flow cytometry) of human monocytes infected with AdV-SOCS1 for 24 h. Infection with three batches of virus used in this study are shown. Batch A (open circles), batch B (closed circles), batch C (open squares). <i>C.</i> SOCS1 protein expression in cell lysates from human monocytes infected with AdV-SOCS1 and AdV-GFP for 24 h.</div>
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<p> FIGURE 2. SOCS1 regulation of TNFα levels… </p>
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<p> FIGURE 2. SOCS1 regulation of TNFα levels 24 h, but not 2 h following LPS-activation… </p>
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FIGURE 2. SOCS1 regulation of TNFα levels 24 h, but not 2 h following LPS-activation of human monocytes
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<div class="figure-caption-contents"><i>A.</i> TNFα levels in culture supernatants 2 h (n=9 donors for all groups except AdV-SOCS3, n=5 donors) and 24 h (n=18 donors for all groups except AdV-SOCS3, n=12 donors) after LPS (500 ng/ml) exposure. <i>B.</i> IL-6 and IL-10 levels assayed 24 h following LPS exposure. Each point represents the mean cytokine level calculated from triplicate cultures of cells from individual donors. * p&lt;0.05, ns = not significant.</div>
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<p> FIGURE 3. Dose-dependent control of TLR4-induced TNFα</p>
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<p> FIGURE 3. Dose-dependent control of TLR4-induced TNFα production by SOCS1 </p>
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<div class="figure-caption-contents"><i> A, B. </i> Increasing the MOI…</div>
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FIGURE 3. Dose-dependent control of TLR4-induced TNFα production by SOCS1
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<div class="figure-caption-contents"><i>A, B.</i> Increasing the MOI of AdV-SOCS1 significantly increased both the percentage of GFP+ cells and the MFI for GFP+ cells. Monocytes from a representative donor were infected with No virus, AdV-GFP (MOI 50), AdV-SOCS3 (MOI 50) or AdV-SOCS1 at MOI 1 to 100 for 24 h prior to analysis of the number of GFP positive cells or MFI by flow cytometry. C. After 24 h, the AdV-infected cells were incubated with LPS (50 or 500 ng/ml) for 24 h and TNFα levels measured in the culture supernatants. Data are shown as mean ± SEM for triplicate cultures. Significant differences between AdV-GFP- and AdV-SOCS1-infected cells are shown by *p&lt;0.05, ** p&lt;0.01. # represents a significant difference between uninfected and AdV -infected cells, p&lt;0.05.</div>
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id="article-image-3"
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<p> FIGURE 4. SOCS1 does not modulate the… </p>
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<p> FIGURE 4. SOCS1 does not modulate the TLR-induced NFκB pathway </p>
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<div class="figure-caption-contents"><i> A. </i> Activation of IκBα by…</div>
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FIGURE 4. SOCS1 does not modulate the TLR-induced NFκB pathway
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<div class="figure-caption-contents"><i>A.</i> Activation of IκBα by phosphorylation on Ser<sup>32</sup> and its subsequent degradation for 2 h following LPS exposure in AdV-GFP- and AdV-SOCS1-infected monocytes. Phospho-IκBα and IκBα levels for cells from 5 donors were quantified and normalized to β-tubulin; mean ± SEM. The kinetics and extent of IκBα-phosphorylation<sup>Ser32</sup> and IκBα-degradation were not significantly different between AdV-GFP- and AdV-SOCS1-infected monocytes. <i>B.</i> NFκB EMSA. Monocytes were left uninfected or infected with AdV-GFP or AdV-SOCS1 for 24 h. Cells were stimulated with or without LPS (500 ng/ml) or Pam<sub>3</sub>CSK4 (300 ng/ml) for 30 min and nuclear extracts prepared. Specificity of binding was determined by incubating nuclear lysates in the presence of a 50 fold excess of a cold NFκB probe (NFκB CP). <i>C.</i> Cumulative results for binding of the NFκB probe by nuclear extracts from 3 donors. The fold change in NFκB DNA binding capacity was determined by densitometry (ns = not significant). <i>D.</i> Nuclear lysates were prepared from CD14+ monocytes from a representative donor incubated for 24 h with no virus, AdV-GFP or AdV-SOCS1 prior to incubation with LPS for 30 min. The complexes binding to the NFκB probe were identified by supershifting with an anti-p65 (+p65) or anti-p50 antibody (+p50). The effect of addition of 50 fold excess of unlabelled NFκB (NFκB CP) or mutant NFκB (mNFκB CP) probe was determined.</div>
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<p> FIGURE 5. Regulation by SOCS1 of AdV-induced… </p>
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<p> FIGURE 5. Regulation by SOCS1 of AdV-induced IFN-dependent pathways </p>
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<div class="figure-caption-contents"><i> A. </i> Relative levels of IFNβ, IRF-1…</div>
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FIGURE 5. Regulation by SOCS1 of AdV-induced IFN-dependent pathways
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<div class="figure-caption-contents"><i>A.</i> Relative levels of IFNβ, IRF-1 and IRF3 mRNA in AdV-GFP- and AdV-SOCS1-infected cells were determined for 3 individual donors using RT<sup>2</sup> Profiler PCR Arrays (SABiosciences). Data from the 3 individual experiments are shown as mean fold change in mRNA expression relative to uninfected or AdV-SOCS1-infected monocytes. The dotted line indicates no change. <i>B.</i> MxA mRNA expression levels were measured by real time PCR and normalized to the expression of the housekeeping gene UBE2D2. <i>C.</i> Inflammatory cytokine mRNA levels, excluding IL-10 and IL-12, were significantly reduced in AdV-SOCS1-infected monocytes. <i>D.</i> Monocytes from a representative donor were left uninfected or infected with AdV-GFP, AdV-SOCS1 or AdV-SOCS3 for 24 h before exposure to LPS for up to 6 h. TNFα mRNA levels were measured by real time PCR and normalized to UBE2D2. <i>E.</i> IFNβ levels were measured in the culture supernatants of triplicate cultures after 6 h exposure to LPS. <i>F.</i> Activation of STAT1. STAT1 phosphorylation<sup>Tyr701</sup>, in response to LPS exposure for 2 h, was determined for cells from a representative donor using Western blotting. Densitometric measures are shown upon normalization to the amount of total STAT1. At time 0, STAT1 activation was elevated in AdV-GFP-infected cells compared to cells incubated with no virus or AdV-SOCS1. After 2 h, STAT1-normalized levels of phosphorylated STAT1 were reduced in cells infected with AdV-SOCS1. All data are represented as mean ± SEM, significant differences are shown as *p&lt;0.05 or ** p &lt;0.01.</div>
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<p> FIGURE 6. SOCS1-regulation of pro-inflammatory cytokine production… </p>
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<p> FIGURE 6. SOCS1-regulation of pro-inflammatory cytokine production by TLR-activated synovial fluid macrophages <i> ex vivo </i> </p>
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<div class="figure-caption-contents"><p> Mononuclear… </p></div>
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FIGURE 6. SOCS1-regulation of pro-inflammatory cytokine production by TLR-activated synovial fluid macrophages <i>ex vivo</i>
</strong>
<div class="figure-caption-contents">Mononuclear cells were isolated from synovial fluid of patients with inflammatory arthritis, incubated overnight in M-CSF, infected for 24 h with no virus, AdV-GFP and AdV-SOCS1 and then exposed to LPS (500 ng/ml) for 24 h. TNFα levels were determined in the culture supernatants, mean ± SEM for triplicate cultures.</div>
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<p> FIGURE 7. Proposed regulation of LPS-induced TNFα</p>
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<p> FIGURE 7. Proposed regulation of LPS-induced TNFα production by SOCS1 </p>
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<div class="figure-caption-contents"><i> A. </i> In AdV-infected macrophages STAT1…</div>
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FIGURE 7. Proposed regulation of LPS-induced TNFα production by SOCS1
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<div class="figure-caption-contents"><i>A.</i> In AdV-infected macrophages STAT1 activation, IFNβ production and MxA mRNA levels were enhanced. In AdV-SOCS1-infected cells relative to AdV-GFP-infected cells there was reduced IRF1 mRNA, IFNβ mRNA, STAT1 phosphorylation and reduced MxA mRNA levels, highlighting the capacity of SOCS1 to control AdV-mediated signaling. <i>B.</i> TLR4 signals via an early MyD88-dependent and a late MyD88-independent, TRIF-dependent pathway. The rapid production of TNFα following receptor activation (+ LPS early) can be attributed to the amplification of signals through the NFκB pathway with NFκB transcriptional activity driving the expression of proinflammatory genes. The sustained production of TNFα, in response to TLR4 activation (+LPS late), reflects the co-operative actions of the MyD88-dependent and MyD88-independent pathways. Recruitment of the adaptors TRIF and TRAM initiate the dimerization and translocation of IRF3 which regulates the production of type I IFN. IFNβ binds to and activates its receptor, IFNAR leading to the activation and nuclear translocation of STAT1 which may drive the expression of both IFN-inducible and pro-inflammatory genes. In addition, TRAF6 recruitment contributes to NFκB activation. Further, cross-talk between the IFN and NFκB pathways is proposed with both known and unknown targets of SOCS1 indicated.</div>
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