Figure 5. Effect of SLRef RNA on HCV replication ex vivo. (A) Schematic representation of HCV monocistronic replicon RNA (adapted from Frese et al. 2003).³³ (B) SLRef, SLRf, SLRe RNAs (500pmol) were transfected in to the Huh7 cells harboring HCV monocistronic replicon construct. 48 h post transfection RNA was isolated and subjected to RT PCR. cDNA synthesized from RT-PCR was then subjected to qRT-PCR for analyzing the relative HCV RNA level. The white bar indicates the HCV mRNA level after transfection with small RNAs. Percentage of RNA level calculated considering untranfceted Huh7 cells harboring HCV monocistronic replicon construct . (C) Curing assay of replicon cells in the presence or absence of SLRef RNA. (D) Schematic representations of HCV JFH1 construct(Adapted from ref. 34) . (E) in vitro transcribed HCV RNA from HCV JFH1 construct was transiently co-transfected with SLRef RNA and SLRf RNA on the Huh7 cells. 48 h post transfection RNA was isolated and then analyzed by qRT-PCR for detecting the HCV RNA level. The bar indicates the HCV mRNA level. Percentage RNA level represents the effect of SLRef RNA on HCV RNA level. (F) Schematic representation of pSGR-JFH1/luc RNA (adapted from ref. 23).(G)pSGR-JFH1/luc RNA was co transfected with SLRef RNA (250pmol and 500pmol) into Huh7 cells and after 24hrs of post transfection RNA was isolated and subjected to RT PCR. cDNA synthesized from RT-PCR was then subjected to quantitative real time PCR for analyzing the relative HCV RNA level and lysate was analyzed for Luciferase activity, which indicates the HCV IRES activity.