FIG. 7.

The replication defect caused by the C-terminal residues of H77 NS4B is completely suppressed by the NS3 mutations Q1112R and S1369R. (A) Schematic representation of NS4B showing the alignment of the 34 C-terminal amino acids of Con1 and H77 NS4B. Divergent residues are shown in boldface, and the arrows indicate the mutations tested. (B and C) At 96 h posttransfection, NS5A protein expression was analyzed by immunoblotting and the number of HCV RNA molecules in 1 μg of total cellular RNA was quantified by real-time RT-PCR. Below the NS5A blots, HCV RNA levels (% RNA) are shown as percentages of the C1/WT level, which has been set at 100%. NS5A protein expression and HCV RNA levels are representative of at least three independent experiments, except for C1/Q235A, C1/I242V, and C1/K247R, which were tested only once.