FIG 6.

Impaired NS2 palmitoylation reduced the colocalization of NS2^YFP with Erlin-2 and Core but not with E2. (Top) Schematic of 2^YFPE3, which has an organization identical to that of 2E3 but with NS2^YFP replacing NS2. (A) Western blot detection of palmitoylated NS2 in cell lysates derived from wt or C113S(TCA) mutated 2^YFPE3 RNA replicating cells following APEGS assay. The PEGylated NS2^YFP is indicated by an asterisk. n.s., nonspecific bands. (B) Extracellular HCV titer determined using culture supernatants collected at different time points postelectroporation of wt or NS2^YFP/C113S(TCA) mutated 2^YFPE3 RNA. (C to E) Confocal imaging (left) and Pearson’s correlation coefficient analysis (right) of 30 different images to detect the colocalization of NS2^YFP with Erlin-2 (C), Core (D), and E2 (E) in cells at day 3 postelectroporation of wt or NS2^YFP/C113S(TCA) mutated 2^YFPE3 RNA. The asterisks indicate statistically significant differences between paired values based on an unpaired Student t test with Welch’s correction. *, P < 0.05; ** P < 0.005. A difference with a P value of >0.05 was considered not significant (ns).