Fig. 7.

Modulation of HCV replication/translation by siRNA-mediated down-regulation of Stau1, ADAR1, DDX6, PA2G4, HSP60, and IGF2BP1. The control MH14 cells carrying replicating HCV replicon were grown for 24 h (lane 1) and then transfected with 20 nm of siRNA duplexes targeting individual cellular factors (lanes 4–6) or with control siRNA duplexes (lane 3). A mock transfection was also done using only transfection reagent (lane 2). Cells were grown for 72 h after transfection, and total protein and RNA were isolated. A shows controls (lanes 1–3) as well as siRNA-mediated down-regulation of targeted host cell proteins (lanes 4–6) as assessed by Western blotting, inhibition, or stimulation of HCV replication by RT-PCR (5′NTR region of the HCV genome) and HCV translation by Western blotting of HCV NS5A. B–D, respectively, show the protein bands of targeted cellular proteins, corresponding RT-PCR of HCV RNA, and expression of HCV NS5A quantified by Quantity One software (Bio-Rad).