CaMKII γ or δ knockdown did not affect NS5A S235 phosphorylation or HCV RNA levels in the infected Huh7.5.1 cells.

<p>(A) Representative and summary (B) of the immunoblotting for NS5A and NS5A phosphorylation at S235 (pS235) in the HCV-infected Huh7.5.1 cells upon CaMKII γ or δ knockdown. One day after the cells were infected with HCV (J6/JFH1), the cells were subjected to small hairpin RNA (shRNA)-mediated CaMKII knockdown for 6 days prior to the immunoblotting analysis. Three and two shRNA sequences were used for CaMKII δ and γ Knockdown. Values are Mean ± SEM (n = 3). Asterisk indicates significance i.e. p<0.05, t-test against the values in the non-targeted control cells (shCon). Protein abundance was quantified with the Li‐Cor scanner, adjusted for the loadings (i.e. against β-actin) and normalized with the values in the control cells. (C and D) CaMKII mRNA and HCV RNA levels measured with quantitative RT-PCR in the HCV-infected Huh7.5.1 cells with or without CaMKII knockdown. The experiments were done as those in a and b except that RNA was collected for the analysis. The GAPDH mRNA levels were analyzed as loading controls. Relative RNA abundance was normalized to the values of the control cells.