FIG 6.

HJV uORF-mediated translational regulation responds to eIF2α phosphorylation in hepatic cells. The WT, uORF1, uORF2, and “no uORFs” constructs were separately cotransfected with a plasmid encoding Renilla luciferase (pRL-TK) into HeLa and HepG2 cells. Six hours after transfection, cells were left untreated (dimethyl sulfoxide [DMSO]) or treated with 1 μM thapsigargin for 24 h. (A) Representative Western blot analyses of HeLa and HepG2 cell extracts left untreated or treated as indicated. Immunoblotting was performed using human eIF2α- and human phosphorylated eIF2α-specific antibodies to control for eIF2α phosphorylation conditions, and a human α-tubulin-specific antibody was used to control for variations in protein loading. (B) Relative luciferase activities were quantified as described in the legend to Fig. 1B.