Figure 4. Inhibition of HCV IRES mediated translation by SLRef RNA. (A) Schematic representation of HCV bicistronic RNA. (B) Small RNAs were co-transfected with HCV bicistronic RNA to the Huh 7 cells. 12h post transfection lysate was collected and analyzed for luciferase activity. FLuc activity represented in the white bar indicates the effect of small RNAs on HCV RNA translation. RLuc activity represented in the black bars indicates the effect of small RNAs on cap-dependent translation. Luciferase activity of control was expressed as 100%. Data from the co-transfection experiments are expressed as means ± SD of four independent replicates. (C) and (D) Representative polysome profile of Rep2a cells after transfecting with pSUPER NSP and pSUPER SLRef. (E) Quantifications of HCV genome in the polysomes and free fraction by qRT-PCR. HCV RNA levels were normalized with GAPDH RNA level.