FIG 2.

Identification of NS5A-associated kinases involved in the HCV life cycle. (A) siRNA-based gene silencing of NS5A-associated kinases. Huh7.5.1 cells were transfected with siRNAs targeting the indicated genes and were harvested 5 days later for immunoblotting (IB) and RT-qPCR to confirm knockdown efficiencies. mRNA levels of target genes relative to GAPDH mRNA were normalized with values for transfection of control siRNAs (siCtrl), which were set at 100%. Results represent the means ± standard deviations from three independent transfections of siRNA. Mock represents transfection without siRNA. (B) Infectious HCV production and cell viability following knockdown of NS5A-associated kinases. Huh7.5.1 cells were infected with JFH-1 virus at an MOI of 1, 2 days after siRNA transfection. Culture supernatants and cells were harvested 3 days later to determine infectious virus yields (upper panel) and cell viability (lower panel), respectively. Cell viability for each transfection was normalized to that for mock transfection (mock), which was set at 100%. Results shown represent the means ± standard deviations from three independent transfections of siRNA. Mock, transfection without siRNA.