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. Author manuscript; available in PMC: 2016 Aug 16.
Published in final edited form as: Sci Signal. 2016 Feb 16;9(415):ra18. doi: 10.1126/scisignal.aac4374

Fig. 5. ONC201 activates the ATF4 pathway through the eIF2α kinases HRI and PKR.

Fig. 5

(A) Western blotting analysis for phosphorylated and total eIF2α was performed in the indicated cell types treated with ONC201. (B to D) Western blotting analysis for the indicated proteins in lysates from HCT116 cells that were transfected with: siRNA against one (B) or two eIF2α kinases (C) as indicated for 24 hours, and subsequently treated with ONC201 (10 μM) for 12 hours. (D) Western blotting analysis for indicated proteins in lysates from HCT116 cells that were transfected with plasmid constructs for wild-type (wt) and a nonphosphorylatable eIF2α mutant (S51A) for 48 hours, and subsequently treated with ONC201 for 12 hours. (E) Western blotting analysis for cyclin D1 in HCT116 cells transfected with the indicated siRNA(s) and treated with ONC201 for 24 hours. Blots in (A) to (E) are representative of at least two independent experiments.