FIG 3.

Restoration of NS5A hyperphosphorylation and infectious virus yields by ectopic expression of siRNA-resistant CKI-α. (A) Huh7.5.1 cells were cotransfected with the indicated siRNAs and plasmid DNAs. The next day, cells were infected with JFH-1 virus at an MOI of 0.5. Culture supernatants and cells were harvested an additional 3 days later for measurement of virus yields and immunoblotting (IB). The p58/p56 ratios were calculated after quantifying the band intensities of NS5A. Values shown represent the means ± standard deviations from three replicate experiments. *, P < 0.05; **, P < 0.01. (B) Huh-7 cells were transfected either with CKI-α siRNA (siCKI-α) or with an irrelevant control siRNA (siCtrl). The next day, cells were retransfected with pJFH1 and pCAG-CKI-α/m6 or empty vector (pCAGGS), followed by infection with vaccinia virus expressing the T7 RNA polymerase at an MOI of 10. NS5A bands were quantified by densitometric analysis, and the p58/p56 ratios were calculated. Immunoblotting images and values shown are representative of two independent experiments.