Figure 6

SLC3A2 is required for HCV propagation. (A) Huh7.5 cells were transfected with 40 nM of the indicated siRNAs for 48 h and then infected with Jc1. At 48 h postinfection, intracellular RNA levels were analyzed by qRT-PCR (upper panel), and protein levels were determined by immunoblot assay using the indicated antibodies (lower panel). (B) Naïve Huh7.5 cells were infected with Jc1 in supernatant harvested from the first infection. At 48 h postinfection, both RNA (upper panel) and protein (lower panel) levels were determined by qRT-PCR and immunoblot analysis, respectively. (C) Huh7.5 cells were transfected with 40 nM of the indicated siRNAs, individually. At 96 h after transfection, cell viability was assessed by WST assay. Negative, universal negative control siRNA; positive, HCV-specific siRNA targeting the 5′ nontranslated region (NTR) of Jc1. (D) Huh7.5 cells were transfected with the indicated siRNAs. At 24 h after transfection, cells were further transfected with either wild-type or siRNA-resistant SLC3A2 mutant plasmid, followed by Jc1 infection. At 48 h postinfection, total cell lysates were immunoblotted with the indicated antibodies. Flag-SLC3A2 WT, Flag-tagged SLC3A2 wild-type; Flag-SLC3A2 Res, Flag-tagged siRNA-resistant SLC3A2 mutant.