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<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="JF7LusxrLlH40NkDsahRdjM027repADi0sRQ7i0NlWuyUeqt5tUTHriEAb9qibbC">
<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="JF7LusxrLlH40NkDsahRdjM027repADi0sRQ7i0NlWuyUeqt5tUTHriEAb9qibbC">
<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="JF7LusxrLlH40NkDsahRdjM027repADi0sRQ7i0NlWuyUeqt5tUTHriEAb9qibbC">
<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="JF7LusxrLlH40NkDsahRdjM027repADi0sRQ7i0NlWuyUeqt5tUTHriEAb9qibbC">
<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="JF7LusxrLlH40NkDsahRdjM027repADi0sRQ7i0NlWuyUeqt5tUTHriEAb9qibbC">
<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="10766741">
<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="http://www.genesdev.org/cgi/pmidlookup?view=long&amp;pmid=10766741"
target="_blank"
rel="noopener"
ref="linksrc=fulltextorjournal_fulltext&amp;is_pmc=False&amp;PrId=3051&amp;itool=Abstract-def&amp;log$=linkouticon&amp;uid=10766741&amp;db=pubmed&amp;nlmid=8711660"
title="See full text options at HighWire"
data-ga-category="full_text"
data-ga-action="HighWire"
data-ga-label="10766741"
><img src="https://cdn.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--highwire.stanford.edu-icons-externalservices-pubmed-genesdev_final_free.gif" alt="HighWire full text link"><span class="text">
HighWire
</span></a><a class="link-item
pmc
"
href="https://pmc.ncbi.nlm.nih.gov/articles/pmid/10766741/"
target="_blank"
rel="noopener"
ref="linksrc=fulltextorjournal_fulltext&amp;is_pmc=True&amp;PrId=3494&amp;itool=Abstract-def&amp;log$=linkouticon&amp;uid=10766741&amp;db=pubmed&amp;nlmid=8711660"
title="Free full text at PubMed Central"
data-ga-category="full_text"
data-ga-action="PMC"
data-ga-label="10766741"
><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="/10766741/citations/"
data-citation-style="nlm"
data-pubmed-format-link="/10766741/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="10766741" 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%2F10766741%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="JF7LusxrLlH40NkDsahRdjM027repADi0sRQ7i0NlWuyUeqt5tUTHriEAb9qibbC"><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="[^&quot;&amp;=&lt;&gt;\/]*" title="The following characters are not allowed in the Name field: &quot;&amp;=&lt;&gt;/"
maxlength="100"
data-ga-category="collections_button"
data-ga-action="create_collection"
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Severe liver degeneration and lack of NF-kappaB activation in NEMO/IKKgamma-deficient mice
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Phosphorylation of IkappaB, an inhibitor of NF-kappaB, is an important step in the activation of the transcription factor NF-kappaB. Phosphorylation is mediated by the IkappaB kinase (IKK) complex, known to contain two catalytic subunits: IKKalpha and IKKbeta. A novel, noncatalytic component of this kinase complex called NEMO (NF-kappaB essential modulator)/IKKgamma was identified recently. We have generated NEMO/IKKgamma-deficient mice by gene targeting. Mutant embryos die at E12.5-E13.0 from severe liver damage due to apoptosis. NEMO/IKKgamma-deficient primary murine embryonic fibroblasts (MEFs) lack detectable NF-kappaB DNA-binding activity in response to TNFalpha, IL-1, LPS, and Poly(IC) and do not show stimulus-dependent IkappaB kinase activity, which correlates with a lack of phosphorylation and degradation of IkappaBalpha. Consistent with these data, mutant MEFs show increased sensitivity to TNFalpha-induced apoptosis. Our data provide in vivo evidence that NEMO/IKKgamma is the first essential, noncatalytic component of the IKK complex.
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<div class="figure-caption-contents"><p> Generation of <i> NEMO/IKKγ </i> <sup> / </sup> mice.… </p></div>
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<p> Figure 1 </p>
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<div class="figure-caption-contents"><p> Generation of <i> NEMO/IKKγ </i> <sup> / </sup> mice. ( <i> A </i> ) Wild-type <i> NEMO/IKKγ </i> locus ( <i> top… </i> </p></div>
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<div class="figure-caption-contents">Generation of <i>NEMO/IKKγ</i><sup>/</sup> mice. (<i>A</i>) Wild-type <i>NEMO/IKKγ</i> locus (<i>top</i>). Exon 1 and 2 are shown as open boxes and the ATG translation start site is indicated. (P) External flanking probe, (B) <i>Bam</i>HI, (RV) <i>Eco</i>RV, (S) <i>Spe</i>I. The targeting vector (<i>middle</i>) was designed to replace exon 1 and exon 2 of the wild-type locus with a neomycin cassette in antisense orientation. Flanking probe P detects a 6-kb <i>Bam</i>HI fragment corresponding to the wild-type allele and a 3-kb <i>Bam</i>HI fragment for the targeted allele (<i>bottom</i>). (<i>B</i>) Genomic Southern blot of <i>Bam</i>HI-digested DNA from E12.5 embryos of the indicated genotypes, showing the 6-kb wild-type band and the 3-kb mutant band detected by probe P. (<i>C</i>) Genotyping by PCR of DNA from E12.5 embryos using the primers described in Material and Methods. (<i>D</i>) Western blot analysis of <i>NEMO/IKKγ</i> protein expression in cytoplasmic extracts of MEFs of the indicated genotypes.</div>
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<div class="figure-caption-contents"><p> Massive liver apoptosis in the… </p></div>
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<div class="figure-caption-contents"><p> Massive liver apoptosis in the absence of <i> NEMO/IKKγ </i> . ( <i> AD </i> ) Histological… </p></div>
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<div class="figure-caption-contents">Massive liver apoptosis in the absence of <i>NEMO/IKKγ</i>. (<i>AD</i>) Histological analysis of liver sections from E12.5 (<i>A,B</i>) and E13.0 (<i>C,D</i>) wild-type (<i>A,C</i>) and <i>NEMO/IKKγ</i>-deficient (<i>B,D</i>) embryos. Hematoxylin and eosin staining; high-power view. (<i>D</i>) The inset shows a low power view of the same region shown in <i>D</i>. The arrow points to a large area of tissue loss in which erythrocytes have accumulated. (<i>E,F</i>) Detection of cell proliferation as determined by BrdU incorporation. Pregnant females were injected IP with BrdU 60-min prior to sacrifice. Sections of E12.5 wild-type and mutant embryos were prepared and immunostained with anti-BrdU antibody. A high power view of the liver is shown. (<i>G,H</i>) Enhanced apoptosis in livers of E12.5 <i>NEMO/IKKγ</i>-deficient embryos. TUNEL assays were performed on sections of wild-type and mutant E12.5 embryos. A high power view of the liver is shown in both cases.</div>
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<div class="figure-caption-contents"><p> Impaired NF-κB activation in <i> NEMO/IKKγ</i> </p></div>
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<div class="figure-caption-contents"><p> Impaired NF-κB activation in <i> NEMO/IKKγ </i> -deficient cells. Wild-type or <i> NEMO/IKKγ </i> <sup> / </sup> MEFs were… </p></div>
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Figure 3
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<div class="figure-caption-contents">Impaired NF-κB activation in <i>NEMO/IKKγ</i>-deficient cells. Wild-type or <i>NEMO/IKKγ</i><sup>/</sup> MEFs were incubated with mouse recombinant TNFα (10 ng/ml) (<i>A</i>), LPS (15 μg/ml) (<i>B</i>), or IL-1 (10 ng/ml) (<i>C</i>) for the indicated times. <i>NF-κB</i> activation in 10 μg of nuclear extract was determined by gel shift assay as described in Materials and Methods.</div>
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<div class="figure-caption-contents"><p> Defective TNFα-induced IκBα phosphorylation and… </p></div>
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<p> Figure 4 </p>
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<div class="figure-caption-contents"><p> Defective TNFα-induced IκBα phosphorylation and degradation and IκB kinase activity in the absence… </p></div>
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Figure 4
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<div class="figure-caption-contents">Defective TNFα-induced IκBα phosphorylation and degradation and IκB kinase activity in the absence of <i>NEMO/IKKγ</i>. (<i>A</i>) IκBα phosphorylation. (<i>Top</i>) Wild-type and <i>NEMO/IKKγ</i>-deficient MEFs were treated with TNFα (10 ng/ml) for the indicated times followed by Western blot analysis of cytoplasmic extracts with anti-phospho-<i>IκBα</i>-specific antibody. (<i>Bottom</i>) Western blot of the same extracts with anti-<i>IκBα</i> antibody as a control to show IκBα protein expression in mutant cells. (<i>B</i>) IκBα degradation. Wild-type (<i>top</i>) and <i>NEMO/IKKγ</i>-deficient (<i>bottom</i>) MEFs were treated with TNFα (10 ng/ml) for the indicated times followed by Western blot analysis with anti-<i>IκBα</i> specific antibody. (<i>C</i>) IκB kinase activity. Wild-type and <i>NEMO/IKKγ</i>-deficient MEFs were either left in medium alone or treated with TNFα (10 ng/ml) for 7 min. Whole cell extracts were immunoprecipitated with anti-<i>IKKα</i> antibody and in vitro kinase activity was determined as described in Materials and Methods using GSTIκBα (172) (<i>left</i>) or GSTIκBα (172) S/A (<i>right</i>) as a substrate. One representative experiment of four is shown.</div>
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<div class="figure-caption-contents"><p> Impaired expression of <i> NF-κB </i> target… </p></div>
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<div class="figure-caption-contents"><p> Impaired expression of <i> NF-κB </i> target genes in <i> NEMO/IKKγ </i> -deficient MEFs. ( <i> A </i> )… </p></div>
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Figure 5
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<div class="figure-caption-contents">Impaired expression of <i>NF-κB</i> target genes in <i>NEMO/IKKγ</i>-deficient MEFs. (<i>A</i>) TNFα-induced <i>IκBα</i> mRNA synthesis. Wild-type and <i>NEMO/IKKγ</i>-deficient MEFs were stimulated with TNFα (10 ng/ml) for the indicated times and 20 μg of RNA was analyzed by Northern blot analysis using a <sup>32</sup>P-labeled random priming probe corresponding to the full-length <i>IκBα</i> cDNA. <i>β-actin</i>, loading control. (<i>B</i>) IL-1 induced IL-6 production. Wild-type and <i>NEMO/IKKγ</i>-deficient MEFs (3×10<sup>4</sup>) were left untreated or stimulated with IL-1 (10 ng/ml) for 16 hr. Levels of IL-6 (pg/ml) in culture supernatants were determined by ELISA. Values shown are the mean and standard deviation of triplicate samples.</div>
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<p> Figure 6 </p>
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<div class="figure-caption-contents"><p> TNFα-induced cytotoxicity. ( <i> A </i> )… </p></div>
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<p> Figure 6 </p>
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<div class="figure-caption-contents"><p> TNFα-induced cytotoxicity. ( <i> A </i> ) TNFα-induced cytotoxicity. Wild-type and <i> NEMO/IKKγ </i> -deficient (2 ×… </p></div>
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Figure 6
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<div class="figure-caption-contents">TNFα-induced cytotoxicity. (<i>A</i>) TNFα-induced cytotoxicity. Wild-type and <i>NEMO/IKKγ</i>-deficient (2×10<sup>5</sup>) MEFs were left untreated or stimulated with the indicated concentrations of TNFα for 18 hr. Cells were then harvested and stained with PI/AnnexinV-FITC. Values shown are the percentage of viable cells after treatment relative to untreated controls. (<i>B</i>) Cytotoxicity induced by TNFα plus cycloheximide. Wild-type and <i>NEMO/IKKγ</i>-deficient MEFs (2×10<sup>5</sup>) were left untreated or stimulated with TNFα (10 ng/ml) in combination with the indicated concentrations of cycloheximide for 18 hr. Cells were harvested and stained as described for <i>A</i>, and values shown were determined as for <i>A</i>.</div>
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<div class="figure-caption-contents"><p> TNFα-induced cytotoxicity. ( <i> A </i> )… </p></div>
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<p> Figure 6 </p>
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<div class="figure-caption-contents"><p> TNFα-induced cytotoxicity. ( <i> A </i> ) TNFα-induced cytotoxicity. Wild-type and <i> NEMO/IKKγ </i> -deficient (2 ×… </p></div>
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Figure 6
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<div class="figure-caption-contents">TNFα-induced cytotoxicity. (<i>A</i>) TNFα-induced cytotoxicity. Wild-type and <i>NEMO/IKKγ</i>-deficient (2×10<sup>5</sup>) MEFs were left untreated or stimulated with the indicated concentrations of TNFα for 18 hr. Cells were then harvested and stained with PI/AnnexinV-FITC. Values shown are the percentage of viable cells after treatment relative to untreated controls. (<i>B</i>) Cytotoxicity induced by TNFα plus cycloheximide. Wild-type and <i>NEMO/IKKγ</i>-deficient MEFs (2×10<sup>5</sup>) were left untreated or stimulated with TNFα (10 ng/ml) in combination with the indicated concentrations of cycloheximide for 18 hr. Cells were harvested and stained as described for <i>A</i>, and values shown were determined as for <i>A</i>.</div>
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