Figure 4

Anti-HCV effects of the IRF-3ER fusion protein in Huh7.5-IRF3ER cells. A. Inhibitory effects of IRF-3ER dimerization on HCV RNA replication. Huh7.5-IRF3ER cells were inoculated with HCV JFH-1 stock at a MOI 0.5 for 14 days and then 4-HT (1 μM) was added at 72, 48, and 24 hours prior to end-point for sample collection. Control indicates the JFH-1 infected Huh7.5-STAT1ER cells without 4-HT treatment for 72 hours. HCV JFH-1 RNA levels were measured by quantitative real-time PCR in triplicate. Relative JFH-1RNA level was calculated as proportion of control (1.0). The data is presented after normalization with an internal GAPDH control. The error bars indicate the variation present in three independent assays. B. Inhibitory effects of IRF-3ER dimerization on HCV IRES-mediated translation. Huh7.5-IRF3ER cells were transfected with the plasmid pRL-HL. After 24 hours of incubation at 37°C and 5% CO₂, transfected Huh7.5-STAT1ER cells were treated with 4-HT for 48, 72, and 96 hours for analyses of luciferase activity. Control Huh7.5-IRF3ER cells received the pRL-HL cDNA plasmid but did not receive 4-HT treatment. Translation efficiency for each sample was calculated as proportion of control (100%). The error bars indicate the variation of three independent assays.