Figure 5.

miR-122 binding does not shield the 5′ terminus of HCV RNA against recognition of IFIT1, while IFIT5 has no effect on HCV RNA accumulation. 3XFLAG-IFIT1, 3XFLAG-IFIT5 and the corresponding empty vector (EV) were transfected into Huh7.5 cells, and two days later, plasmids plus (A, B) FL WT HCV or GNN or (C, D) S1+S2p3 SGR and the replication incompetent S1+S2p3 GND viral RNA and miRNAs were transfected again. Replication was measured by evaluating luciferase production at the indicated timepoints. (E) Western blot was used to confirm expression of IFITs 1 and 5 using antibodies against FLAG and β-actin. (F) Northern blot analysis of FL HCV RNA accumulation during IFIT1 and IFIT5 overexpression at day 2. (G) Densitometry quantification of northern blot analysis in (F) normalized to siCon. All data are representative of at least three independent experiments and statistical significance was determined by paired parametric t test.