Fig 3.

ADAP is critical for efficient cyclin E and Cdk2 accumulation. (A and B) Purified naïve control (Ctrl) or ADAP^−/− CD4 T cells were activated with anti-CD3 and anti-CD28 antibodies for the indicated times, and cell extracts were prepared for qRT-PCR or Western blotting. (A) qRT-PCR showing fold change relative to β-actin for the early activation marker CD69, cyclin E, and Cdk2. (B) Total cell lysates from the indicated time course performed in parallel with the experiments whose results are shown in panel A were subjected to Western blotting with antibodies to cyclin E, Cdk2, and β-actin. (C) Wild-type or ADAP^−/− T cells were activated with anti-CD3 and anti-CD28 antibodies for 24 h. Cyclin E immunoprecipitates or control IgG were prepared, and Western blots were probed for cyclin E and ubiquitin. IP, immunoprecipitation; KO, knockout. Results are representative of two independent experiments performed.