FIG 8.

An increase of CLDN1 localization at the cell surface is not sufficient to increase HCV entry upon either CD81 overexpression or EGF stimulation. (A) Huh-7 cells were transfected with pcDNA, HA–5-HT6, or CD81-YFP alone or in combination, as indicated. At 48 h posttransfection, cells were infected with HCVcc JFH1. At 30 h postinfection, cells were fixed and the infection rate was determined by IFA. (B) After 2 h of starvation, Huh-7 cells were kept nonstimulated or stimulated for 1 h with EGF (1 μg/ml). Western blotting was performed to verify the activation of EGFR through phosphorylation. β-Tubulin was used as a loading control. (C) Huh-7 cells transfected for 48 h with pcDNA, HA–5-HT6, or CD81-YFP, alone or in combination, after 2 h of starvation were treated for 1 h with EGF (1 μg/ml) and infected for 2 h with HCVcc JFH1 concomitantly with EGF (1 μg/ml). At 30 h postinfection, cells were fixed and the infection rate was determined by IFA. Results are presented as means ± SEM (n = 2 [A and C]). One-way (A) or two-way (C) ANOVA followed by the Dunnett or Bonferroni posttest was performed for statistical analysis. ns, nonsignificant.