Huh-7.5 Cell HCV Infection and Characterization

Growth and infection of Huh-7.5 cells with HCV were carried out as described in^{24 (link)}. Briefly, cells were infected with the HJ3-5 chimeric virus at an MOI of 0.1–0.5 and passaged for two weeks until approximately 100% of the cells were HCV positive, as previously described^{24 (link)}. High infection levels were maintained for two weeks by adding 2% human serum to the culture medium, as previously described. HCV-infected and uninfected cells were washed twice with trypsin EDTA Solution B (Biological Industries), spread onto a polylysine adhesion microscope slide (Electron Microscopy Sciences), and incubated for two hours at 37 °C. The cells were fixed in methyl alcohol/acetone for 10 min at room temperature. The slides were then stained with mounting medium, including DAPI (Sigma-Aldrich), and covered with a coverslip (Kaltek).