Figure 2

Analysis of mode-of-action of S. maurus palmatus venom . Huh7.5 cells were infected with HCV and treated with S. maurus palmatus venom (30 μg/ml) at different time points as indicated (A, B) or left untreated as a control. (A) Amounts of HCV infectious particles in the supernatants. Data represent means ± SEM of the data obtained from two independent experiments. *, P <0.05; †, P <0.001, compared with the untreated control; §, below the detection limit; dpi, days post-infection. (B) HCV NS3 protein accumulation in the cells. Virus-infected cells were subjected to immunoblot analysis using monoclonal antibody against the HCV NS3 protein at 1 and 2 days post-infection. GAPDH served as an internal control to verify equal amounts of sample loading. Signal intensities of NS3 were normalized to the corresponding GAPDH signal. (C) Amounts of HCV RNA in the cells. The venom treatment was done only during the post-inoculation (+2 hr) step. HCV RNA amounts were normalized to GAPDH mRNA expression. (D) Amounts of HCV infectious particles inside the cells. The venom treatment was done only during the post-inoculation (+2 hr) step. Virus-infected cells were subjected to 3 cycles of freezing and thawing at −80°C and 37°C, respectively, and HCV infectivity inside the cells was measured.