Figure 4.

10j is required for late stage of HCV lifecycle. (A) Schematic representation of the time-of-addition experiment. Huh-7.5 cells were pre-treated for 2 h before HCVcc ((a) JFH1 virus expressing an NS5A-GFP) infection with 10j at different concentrations or with 1% DMSO (solvent of compound), heparin (2 μg/mL), or sofosbuvir (10 μM) ((a) pre-treatment), or (b) co-treatment with HCVcc, or post treatment at 2 h after HCVcc infection ((c) post-treatment). (B) At 72 h post infection, HCVcc infection was measured by counting the number of GFP positive cells. p-Values are indicated by asterisks (** p < 0.01); Not significant (n.s.). (C) Determination of antiviral activity by dose response curve analysis. (D) Huh7.5 cells were infected with HCVcc and incubated with increasing concentrations of 10j for 72 h. Expressions of NS5A-GFP proteins were quantitated by western blot analysis using a GFP antibody.