Fig 1.

Impaired proliferation and block in G₁-S transition in ADAP^−/− T cells upon CD3/CD28 stimulation. Purified naïve hCAR control (Ctrl) or hCAR/ADAP^−/− T cells were transduced with control adenovirus expressing Thy1.1 (Thy) or adenovirus expressing Thy1.1 and wild-type ADAP (WT), ADAPΔCAR mutant (ΔCAR), ADAPΔTAK mutant (ΔTAK), or ADAPΔSKAP mutant (ΔSKAP). (A) Cells were stained with anti-Thy1.1-APC and analyzed by flow cytometry. (B) Cell lysates were prepared, and Western blotting was performed with anti-ADAP antibody. (C) Cells were labeled with CFSE and stimulated with anti-CD3 and anti-CD28 antibodies for 42 h prior to flow cytometry analysis. (D) Cell cycle analysis was performed on the transduced T cells using DAPI staining. Results for both the proliferation and cell cycle analysis are representative of at least five independent experiments.