FIG. 6.

Colony-forming abilities and 6367 resistance of AB12 and BB7 replicon RNAs carrying escape mutations. Escape mutations that were cloned into AB12 and BB7 replicon cDNAs are shown on the left. Boldface characters indicate the nucleotide mutations that led to amino acid changes. Replicon RNA was coelecroporated into Huh-7 cells with nonsilencing siRNA 6188 mm (NS) or with siRNA 6367 to assess colony growth and 6367 resistance phenotypes. The growth phenotypes for each mutant replicon after coelectroporation with the nonsilencing siRNA 6188 mm are shown in columns AB12+NS and BB7+NS. The growth phenotype for each replicon is expressed as +++ for the wild-type replicon RNA and for mutant replicons that were able to produce colony numbers within 2 times the wild-type levels. Replicon growth was graded as ++ or + if colony counts were less than 2 times or less than 10 times wild-type levels, respectively. The colony numbers for two independent experiments are given in parentheses. The data from the two independent experiments are presented separately due to variability in HCV replicon colony growth in different passages of Huh-7 cells. The relative percent resistance of colony growth following coelectroporation with siRNA 6367 was calculated for each experiment, and the average and standard error for each mutant replicon are shown in the columns labeled AB12 + 6367 and BB7 + 6367. (B) The sequence of the 6367 siRNA duplex is shown.