Fig. 6.
3Cpro does not interact with PKR and the protease activity is not essential for 3Cpro to induce PKR reduction. (A) HEK293T cells grown in the 10-cm dish were co-transfected with Myc-PKR (5 μg) and vector, Flag-L or Flag-3C plasmids (5 μg) for 36 h. The cell lysates were immunoprecipitated with mouse anti-Myc antibody and subjected to western blotting. (B) HEK293T cells grown in the 10-cm dish were co-transfected with Myc-PKR (5 μg) and vector, Flag-L or Flag-3C plasmids (5 μg) for 36 h. The cell lysates were immunoprecipitated with rabbit anti-Flag antibody and subjected to western blotting. (C) HEK293T cells were co-transfected with 2 μg of Myc-PKR plasmids and 3 μg of vector plasmids or various 3 C mutant plasmids for 36 h. The expression of Myc-PKR and host protein NEMO was detected by western blotting. Relative level of Myc-PKR was indicated in folds below the images after normalization with β-actin.