Figure 6.
Inhibitors of PKR result in translational recovery. (A) HeLa S10 cell lysates containing the protein translation machinery, exogenous PKR (10 nM), and 5'-capped luciferase mRNA (50 nM) are incubated for 1 hour at 30 °C in the presence (grey) or absence (black) of HCV-IRES activator dsRNA (50 nM) and increasing amounts of VAI inhibitor (0−2 μM). Measurements were performed in triplicate and standardized relative to maximum Luciferase activity. (B) Western blot probed with an anti-(P-Ser51)-eIF2a antibody (ab4837−50, abcam) for selected reactions shown in (A).