Figure 1
From: cJun N-terminal kinase (JNK) phosphorylation of serine 36 is critical for p66Shc activation
Inhibition of JNK kinases prevents p66ShcS36 phosphorylation under oxidative stress.
MEFs were treated with H2O2 (a) (1 mM, 30 min) or exposed to HR (hypoxia 90 min, reoxygenation 15 min) (b). Cell lysates were probed with the antibodies indicated to assess p66Shc phosphorylation at S36. Representative Western blots (individual experiment performed under the same experimental conditions and run on the same gel) and summary graphs from at least three independent experiments are shown. Signal intensity of control samples was arbitrarily set at 1. Mitochondrial ROS production was measured in p66Shc+/+ and p66Shc−/− MEFs after prooxidant treatment as described in Material and Methods (n ≥ 4) (c) and phospho γH2AX was detected by immunoblotting (d, lower panel) and summary graph is shown in above panel. SP: SP600125, JNK inhibitor and BR: BR796, p38 inhibitor were applied 1 h prior to stress application. Original blot shown in (b) have been cropped and the full length blots are shown in the Supplementary Figure S1b Statistical analysis was performed using t-test or ANOVA (*p < 0.05 **p < 0.01, ***p < 0.001).
