Figure 9.

EWI‐2wint does not inhibit cell‐to‐cell transmission.

A. Huh‐7 donor cells were infected with HCVcc and stained with CMFDA. Acceptor cells are Huh‐7 clones expressing EWI‐2, EWI‐2wint, LAL or Qcc. Huh‐7w7 cells serve as negative control. Co‐culture of donor and acceptor cells with or without neutralizing 3‐11 anti‐E2 mAb allowed to monitor either cell‐to‐cell or total (cell‐to‐cell and cell‐free) transmission of HCV. Cells were labelled with anti‐NS5 mAb followed by PE‐conjugated secondary antibody and analysed by flow cytometry. In these conditions, newly infected cells are negative for CMFDA staining and positive for PE staining. For cell‐free (light grey) and cell‐to‐cell transmission (black), results are presented as percentages relative to the total transmission. Levels of infectivity of each clone are shown in dark grey.

B. Three conditions of cell density (high, medium and low) were used for HCVcc infection, for each Huh‐7 clone. After 48 h, cells were labelled with anti‐NS5 mAb followed by PE‐labelled secondary antibody and analysed by flow cytometry. Results are presented as related percentages to the infection of pcDNA3.1 control cells in high‐confluency condition.

Results are reported as the mean ± SD of three independent experiments (A and B).