Research Paper Volume 2, Issue 10 pp 669—677

Figure 2. Multiple eIF2α kinases are responsible for the induction of eIF2α phosphorylation upon treatment with vorinostat. (A) The indicated MEFs were treated with DMSO (con) or 10 μM vorinostat for the indicated time periods. Protein extracts (50 μg) were subjected to western blot analysis for phosphorylated eIF2α (panels a, c, e, g) and total eIF2α (panels b, d, f, h). (B) GCN2 ^-/- PERK ^-/- MEFs (DKO) were treated together with their isogenic control (WT) with DMSO (con) or 10 μM vorinostat (SAHA) for the indicated time periods. Protein extracts (50 μg) were subjected to western blot analysis for phosphorylated eIF2α (panel a) and total eIF2α (panel b). Representative blots are shown. The ratio of the phosphorylated protein to total normalized to its control is indicated. Quantification of the bands was performed by densitometry using the Scion Image software.