FIG 2.

HCV entry into DGAT1-silenced Huh-7.5 cell lines is impaired. (A and B) DGAT1-silenced cell lines were inoculated with JFH-1 HCVcc at a multiplicity of infection (MOI) of 0.1. After 72 h, infectious HCV virions were quantified in culture supernatants by a colorimetric focus-forming assay (19). The data are presented as focus-forming units (FFU) per ml of culture supernatant (n = 3) (A). Intracellular HCV RNA levels were determined by real-time quantitative PCR and standardized to β-actin mRNA levels (n = 3) (B). (C) DGAT1-silenced cell lines were inoculated with JFH-1 HCVcc at an MOI of 0.5. After 60 h, intracellular HCV core proteins were detected by flow cytometry. Data are representative of two independent experiments. (D and E) DGAT1-silenced cell lines were transfected with 5 μg in vitro-transcribed JFH1 RNA. After 72 h, infectious HCV virions were quantified in culture supernatants by a colorimetric focus-forming assay (19). The data are presented as focus-forming units (FFU) per ml of culture supernatant (n = 3) (D). Intracellular HCV RNA levels were determined by real-time quantitative PCR and standardized to β-actin mRNA levels (n = 3) (E). (F) HCV entry into DGAT1-silenced cell lines was assessed using HCVpp harboring E1 and E2 glycoproteins of various HCV genotypes. Virus pseudoparticles harboring the vesicular stomatitis virus G (VSV-G) envelope glycoprotein were used as a positive control. HCVpp entry was determined by luciferase activity. Data are expressed as percentages of the shRNA-control cell line (n = 3). Bar graphs represent means ± SEM. *, P < 0.001 compared to the control. N.D., not detected.