Fig. 5. dsRNA is the HCV PAMP that activates TLR3 and subsequent chemokine expression.

(A) Gel electrophoresis of in vitro synthesized HCV ssRNAs and dsRNAs derived from the indicated HCV genome region. +ss: positive-sense strand; −ss: negative-sense strand; ds: double-strand RNA duplex; M: GeneRuler 1kb plus DNA ladder (Fermentas). (B) Huh7.5, 7.5-TLR3 and PH5CH8 cells in 24-well plates were mock-treated, incubated in culture medium (5 μg/0.5 ml) or transfected (1 μg/well) with the +ss, −ss or the ds NS-3′NTR RNA for 6 h. Induction of RANTES mRNA was determined by qPCR. (C) Cells were mock-treated, incubated in culture medium or transfected with the indicated HCV dsRNA, or incubated with poly-I:C in culture medium (pIC) for 6 h. Induction of RANTES mRNA was determined by qPCR.