Fig 6.

No CCCM HCV transfer from Huh7.5.1 cells to cells of other hepatoma and nonhepatoma cell lines. HepG2 cells were transfected with pcDNA3-CD81, NKNT3 cells were transfected with pCMV-CLDN1, and 293T cells were transfected simultaneously with both pcDNA3-SR-B1 and pCMV-CLDN. At 48 h posttransfection, the transfected cells were collected for analysis of CD81 expression in HepG2 cells by flow cytometry (a), CLDN1 expression in NKNT3 cells by Western blotting (c), or SR-B1 and CLDN1 expression in 293T cells by Western blotting (e). Simultaneously, these receptor-expressing HepG2, NKNT3, or 293T cells were labeled with PKH26 or CMFDA and cocultured with JFH1-infected Huh7.5.1 cells at a 1:1 ratio for 20 h. The mixed cultures were immunostained for HCV core protein and analyzed by flow cytometry for target cells with CCCM HCV transfer (b, d, and f). Naive Huh7.5.1 cells and pcDNA3-transfected 293T, HepG2, and NKNT3 cells were included as controls in these experiments. hCD81, human CD81; hSR-B1, human SR-B1; NS, not significant.