Fig. 3.

STAT3 regulates ICD marker eIF2α phosphorylation through interaction with PKR. (A) Western blot analysis of STAT3, p‐STAT3^Tyr705, PKR, p‐PKR^Thr446, eIF2α, and p‐eIF2α^Ser51 expression in Huh7 and HepG2.2.15 cells infected with lentiviral STAT3‐shRNA vector or lentiviral vector. (B) Western blot analysis of STAT3, p‐STAT3^Tyr705, PKR, p‐PKR^Thr446, eIF2α, and p‐eIF2α^Ser51 expression in Huh7 and HepG2.2.15 cells stimulated with IL‐6 at the indicated concentrations for 2 h. (C) Western blot analysis of STAT3, p‐STAT3^Tyr705, PKR, p‐PKR^Thr446, eIF2α, and p‐eIF2α^Ser51 expression in Huh7 and HepG2.2.15 cells infected with indicated lentiviral vectors after stimulation with IL‐6. (D) Coimmunoprecipitation with STAT3 antibody. Huh7 and HepG2.2.15 cell extracts were incubated with anti‐STAT3 antibody and Protein A/G Magnetic Beads. Following incubation, PKR was detected by western blot to verify the protein binding. (E) GST pull‐down assay of the interaction between STAT3 and PKR. Western blotting analysis of PKR binding to purified GST or GST‐STAT3 fusion protein using PKR antibody (top). GST or GST‐STAT3 fusion protein was visualised by Coomassie blue staining (bottom). N = 3. Shown is one representative of at least three independent experiments.