FIG 3.

pDC cocultured with hepatoma cells transfected with Jc1 or JFH1 mount IFN-α responses. (A) Huh7.5 cells were transfected with JFH1 or Jc1 RNA and incubated for 24 h. Transfected hepatoma cells were cocultured with pDC for 18 h, and IFN-α was determined in the supernatant by an ELISA method (pDC, n = 4). ***, P = 0.0002; **, P = 0.0072 (Wilcoxon test). (B) Lunet N cells, which are CD81 deficient and thus cannot be infected by HCV, were transfected with JFH1 or Jc1 RNA and incubated for 24 h. Shown is immunofluorescence analysis of JFH1- or Jc1-transfected Lunet N cells stained for NS3. Transfectants were cocultured with pDC for 18 h, and IFN-α was assessed in the supernatant by an ELISA method (pDC, n = 19). ***, P < 0.0001 (Wilcoxon test). (C) Lunet N cells were transfected 3 h, 6 h, 12 h, and 24 h prior to coculture with pDC for 18 h. The TCID₅₀ was determined as a measure of the amount of virus produced during the 18 h of coincubation (pDC, n = 3). The error bars indicate SEM.