Figure 1. Robust Z scores of 2 replicates from the primary whole genome siRNA screen in Huh7/pRepFeo cells.

Peginterferon α-2b efficiently inhibits HCV replication in Huh7/Rep-Feo (A) and OR6 replicon cells (B) in a dosage-dependent manner. Huh7/Rep-Feo (2000/well) or OR6 cells (3000/well) were seeded in a 384-well or 96-well microplate. After 72 hours incubation, the cells were treated with different dosage of Peginterferon α-2b (PEG-IFNα-2b) for an additional 24 hours. Firefly (or Renilla) luciferase activity and cell viability were detected simultaneously. Five IU/mL and 30 IU/mL PEG-IFNα-2b inhibited nearly 80% of the HCV replication in Huh7/Rep-Feo and the OR6 replicon cells, respectively. Small interfering RNA (siRNA) targeting IFNAR1 rescued HCV replication in the presence of PEG-IFNα-2b in Huh7/Rep-Feo (C) and OR6 replicon cells (D). siRNAs were reverse transfected into Huh7/Rep-Feo (2000/well in a 384-well plate) and OR6 cells (3000/well in 96-well plate) at 50nM final concentration. After 72 hours incubation, cells were treated with or without PEG-IFNα-2b (5 IU/mL for Huh7/Rep-Feo, 30 IU/mL for OR6 cell) for an extra 24 hours. Silencing IFNAR1 induced at least 2 fold higher rescue effect over non-targeting siRNA in both replicon systems. Luminescence activity and cell viability were obtained from quadruplicate assays and expressed as mean ± SD. (E) Distribution of robust Z scores of host genes from primary screening. Each point corresponds to the robust Z score of the same siRNA pool in plate A and B.