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

Figure 7.

Figure 7

CIS interacts with activated PKCθ in activated normal T cells, and expression of CIS slightly modulates IL-2–induced STAT5 activation in activated T cells. (a) CD4 T cells from spleen of normal mice were incubated without (lane 1) or with anti-CD3 (lane 2) for 16 h. The activated CD4 T cells were lysed, and cleared lysates were precipitated with anti-CIS antiserum. The precipitates were separated on 10% SDS-PAGE and sequentially blotted by anti-PKCθ and anti-CIS antibodies, respectively. (b) The activation of STAT5 was directly analyzed in CD4 T cells after 15 min incubation with 5 mg/ml anti-CD3 and in CD4 T cells that were prestimulated for 48 h with 5 μg/ml anti-CD3 and then incubated for 15 min with 200 U/ml IL-2. The STAT5 was precipitated with anti-STAT5 antiserum and blotted by an antibody specific for phosphorylated STAT5. Western blot with anti-STAT5 antiserum showed similar amounts of STAT5 in all samples tested.