Figure 1

HCV -3a Core specific siRNAs inhibit Core expression. A) Dose dependent silencing effect of synthetic siRNA against Core gene of HCV-3a. Huh-7 cells were transfected with 0.4 μg of constructed HCV Core vector or Mock along with or without 10, 20 and 40 nM of siRNAs for 24 and 48 hrs. Cells were harvested and relative RNA determinations were carried out using semi-quantitative RT-PCR. Gene expression results are given for increasing concentrations of Csi16, Csi27, Csi151, Csi352, and Csi476 siRNAs against HCV-3a Core. Expression levels for Mock-transfected (M), HCV-3a Core expression plasmid (C), scramble siRNA (Sc), 100 bp DNA Ladder (L) and GAPDH are also shown. B) Quantitative Real Time PCR analysis of Core 3a or Mock-treated Huh-7 cells along with or without 40 nM of siRNAs for 24 and 48 hrs in comparison to Mock. Gene expression results from Real Time PCR shows that Csi16, Csi352, and Csi476 siRNAs against HCV-3a Core decrease RNA expression after 24 and 48 hrs transfection. GAPDH was used as internal control. Each independent experiment was performed having triplicate samples. The p values indicate significant differences between the connected groups. Error bars indicate mean S.D, Csi476 verses other siRNA: p* for 24 and p^ for 48 hrs C). Silencing of HCV-3a Core gene by siRNAs using specific antibodies showed reduction at protein expression level. The protein expression levels were determined by western blot analysis after 24 and 48 hrs transfection with Mock (M), HCV-3a Core expression plasmid (C) with and without HCV-3a siRNAs (Csi16, Csi27, Csi151, Csi352, and Csi476) and scramble siRNA (Sc) in Huh-7 cells.