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. 2000 Mar 20;191(6):985–994. doi: 10.1084/jem.191.6.985

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© 2000 The Rockefeller University Press

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/).

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CIS was induced by TCR activation. (a) Differentially displayed CIS transcript in anti-CD3–stimulated CD4 T cells. (b) Northern analysis of mRNA from CD4 T cells treated with anti-CD3 for different periods of time showing induced expression of CIS (lanes 1–4), and dependence on TCR activation (lane 5). The effect of cycloheximide or cyclosporin A on anti-CD3–induced CIS expression was analyzed by incubation with cyclosporin A together with anti-CD3 or addition of cycloheximide after anti-CD2 stimulation for 6 h (lanes 6–8). Probe specific for G3PDH confirmed that equal amounts of RNA were present in each sample. (c) The expression of CIS in CD4 T cells was analyzed by immunostaining with anti-CIS antibody (Santa Cruz Biotechnology) as described (reference 41). Induced CIS expression was detected in activated T blasts after 20-h stimulation with anti-CD3. Expressed CIS was degraded in 16 h after withdrawal of anti-CD3. (d) RT-PCR analysis of mRNA from CD4 T cells activated with anti-CD3 for different periods of time showing the kinetics of CIS, SOCS2, and IL-2 induction. Primers specific for G3PDH confirmed that equal amounts of RNA were present in each sample.