Research Paper Volume 2, Issue 10 pp 669—677

HDAC pharmacological inhibition promotes cell death through the eIF2α kinases PKR and GCN2

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Figure 1. eIF2αphosphorylation is induced in human cancer and mouse cells upon treatment with vorinostat. (A) A549, HT1080 and HepG2 cells were treated with DMSO (con) or 10 μM of vorinostat (SAHA) for the indicated time periods. Protein extracts (50 μg) were subjected to western blot analysis for phosphorylated eIF2α (panels a, c, e) and total eIF2α (panels b, d, f). (B) eIF2αS/S and eIF2αA/A MEFs were treated with DMSO (con) or 10 μM of 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.