Fig 8. Activation of PKR and phosphorylation of eIF2α phosphorylation in response to transfection of PV subgenomic replicon RNA.

Western blot analysis of p-PKR (T446) (panel A), PKR (panel B), p-eIF2α (S51) (panel C), and eIF2α (panel D) in HeLa cell lysates. Cells were transfected individually with PV subgenomic replicon RNAs: ΔIRES, 3B STOP, and WT or with WT in the presence of 3 mM GuHCl (WT_GuHCl). Six hours posttransfection, cells were processed for western blot analysis and probed using anti-p-PKR (T446), PKR, p-eIF2α (S51), and eIF2α antibodies; α/β tubulin was used as a loading control. Blots provided in Supporting information (S1 Raw Images). GuHCl, guanidine hydrochloride; PKR, RNA-activated protein kinase; PV, poliovirus; T446, threonine-446; WT, wild-type.