HCV 3a 5'UTR specific siRNAs inhibit mRNA expression. A) Huh-7 cells were transfected with 0.4 μg of constructed HCV vector or mock-treated along with or without 10 nM, 20 nM and 40 nM of siRNAs for 24 and 48 hrs. Cells were harvested and relative RNA determinations were carried out using semi-quantitative PCR. Gene expression results from semi-quantitative PCR are given for increasing concentrations of Usi170, Usi212 and Usi272 siRNAs against HCV 3a 5'UTR. Expression levels for mock-transfected (M), HCV 3a 5'UTR expression plasmid (U), scramble siRNA (Sc), 100 bp DNA Ladder (L) and GAPDH are also shown. B) Huh-7 cells were transfected pCR3.1/GFP/5'UTR vector or mock-treated along with or without 40 nM of siRNAs for 24 and 48 hrs. Total cellular RNA extracted, after 24 and 48 hrs post transfection, was quantified by Real Time PCR using gene specific primers in comparison to Mock. Gene expression results from Real Time PCR shows that Usi170 and Usi272 siRNAs against HCV 3a 5'UTR decrease RNA expression both after 24 and 48 hrs post transfection. GAPDH was used as internal control. Three independent experiments were performed having triplicate samples. Error bars indicate mean S.D, p < 0.01.