Figure 8.

Bioflavonoid effects on IRES-mediated translation. A. Schematic of the bicistronic reporter construct used to measure IRES-mediated translation. Renilla luciferase (RLuc) and Firefly luciferase (FLuc) are driven by a 5′ cap and the HCV IRES, respectively. Firefly to Renilla ratios reflect changes in IRES-mediated translation. B. Bioflavonoid effects on IRES-mediated translation. 293T Cells were transfected with the IRES reporter construct and either NS5A or GFP. 12 hours later, cells were treated with 125μM bioflavonoid, followed by luciferase assays 72 hours later. C. Western analysis of the effect of bioflavonoids on HSP70 levels after heat shock. Huh-7.5 cells were treated with 125μM bioflavonoids for 2 hours and subjected to heat shock at 42°C for 30 minutes. After 6 hours of recovery at 37°C, cells were lysed, and Western analysis was performed with antibody against HSP70 (HSPA1A). D. Densitometry of the Western blot in panel C. HSP70 quantities were normalized to tubulin. (*, **, and *** indicate P<0.05, P<0.005, and P<0.0005, respectively. Error bars reflect standard deviation.)