Figure 2. ATM phosphorylation of Mdm2 Ser394 promotes p53-dependent apoptosis after DNA damage.
(A) Ex vivo thymocytes were either left untreated or irradiated with 2Gy for 12 hours and apopototic cells were quantified by flow cytometry (n=3 per genotype). Standard deviation indicated by error bars. Asterisk (p=0.0001) and double asterisk (p=0.0001) indicate significant differences.
(B) Ex vivo thymocytes were either untreated or irradiated with 2Gy for 1 hour, and all cells were then incubated with 12 µM Nutlin-3a for 1 hour. Protein levels were analyzed by western blot.
(C) Ex vivo thymocytes were either left completely untreated or irradiated with 2Gy for 12 hours and incubated with 12 µM Nutlin-3a 3 hours after IR (9 hours total) (n=3 per genotype). Apopototic cells were quantified by flow cytometry Standard deviation indicated by error bars.
(D) Ex vivo thymocytes were preincubated with 10 µM KU55933 for 1 hour. Cells were then either left untreated or irradiated with 2Gy for 12 hours and apopototic cells were quantified by flow cytometry (n=3 per genotype). Standard deviation indicated by error bars.
(E) Ex vivo thymocytes were either left untreated or preincubated with 10 µM KU55933 for 1 hour. Cells were then either untreated or irradiated with 2Gy for 2 hours, and protein levels were analyzed by western blot. See also Figure S2.
