In vitro interaction of HCV E1E2 heterodimers with CD81 and expression of cell markers.

1The ratio [CD81 band intensity in E1E2 IP in Bj/EDTA buffer]/[CD81 band intensity in E1E2 IP in TX/EDTA buffer] was used to evaluate the E1E2/CD81 interaction: -, ≤0.1; +/−, >0.1–0.72Expression of SR-BI and Claudin-1 detected by WB with Cla-1 mAb and JAY.8 polyclonal Ab, respectively. Amount of each protein was compared to Huh-7 cells. The ratio [protein band intensity of the cell line]/[protein band intensity of Huh-7 cell line] was used: -, 3Cell surface expression of CD81 and EWI-2 proteins detected by flow cytometry with 5A6 and 8A12 mAbs, respectively. Analyses gave mean fluorescence intensities (MFI) in the range of 3–6 for negative controls and intensities over 10 for specific stainings.4Expression detected by co-immunoprecipitation with CD81 as described in Figure 1B. The ratio [EWI-2wint band intensity]/[CD81 band intensity] was used to evaluate the expression of EWI-2wint: -, ≤0.01; +/−, >0.01–0.1≤; +, >0.1