Figure 4
From: CBAP promotes thymocyte negative selection by facilitating T-cell receptor proximal signaling
Reduced TCR-induced BIM accumulation in CBAP-deficient thymocytes. (a) Eight-week-old littermates were injected with 20 μg anti-CD3 Ab. Total thymocytes were harvested without treatment or 24 h post injection. BIM and heat shock protein 70 (HSP70) were detected in cell lysates by western blotting. BIM expression in each group was quantified relative to HSP70 and plotted as mean±S.D. (n=4). (b) Thymocytes were cultured on plates coated with anti-CD3 (10 μg/ml) and anti-CD28 (20 μg/ml) Abs. After the indicated times, cells were collected, and BIM and actin were detected by western blotting. Expression of BIM was quantified relative to actin (n=4). (c) Thymocytes from indicated genotypes were cultured onto plates coated with anti-CD3 (1 μg/ml) and anti-CD28 (5 μg/ml) Abs or uncoated plates. After 24 h, thymocytes were collected and stained for CD8, CD4, annexin V, and PI. Survival of DP thymocytes was assessed by flow cytometry with statistical analysis. Specific TCR-induced apoptosis (▵) was calculated as the percentage of dead DP thymocytes in test cultures minus that in untreated cultures. Results were plotted as mean±S.D. (n=4)
