Skip to main content
. 2013 Apr 11;9(4):e1003302. doi: 10.1371/journal.ppat.1003302

Figure 2. TIP47 is required for HCV RNA replication.

Figure 2

A) Schematic representation of HCV Replicon (Con1, genotype 1b), a subgenomic bicistronic HCV replicon with neomycin as a reporter gene, and Luc-JFH1 (genotype 2a), a full-length bicistronic HCV infectious clone with firefly luciferase as a reporter gene. B) Crystal violet-stained Huh7.5 cells transfected with control shRNA or shRNA against TIP47 and HCV replicon RNA (A). G418 was added to the medium (1 mg/ml). Cells were kept under selective pressure for 4 weeks. Only cells actively replicating HCV RNA, and thus expressing the neomycin phosphotransferase survive. C) Quantification of colony numbers/plate in (B) (mean ± standard deviation; n = 3, *P<0.05). D) Western blot analysis of TIP47 expression in shRNA-expressing Huh7.5 cells at the time of transfection in (B). E) Transfection of shRNA-expressing Huh7.5 cells with Luc-JFH1 RNA (A) and Myc-TIP47 expression plasmid (where indicated). HCV replication was measured by determining the fold increase of luciferase (RLU) per well over a 4 h time point (mean ± standard deviation; n = 3; *P<0.05, **P<0.01). F) Western blot analysis of TIP47 expression in shRNA-expressing Huh7.5 cells at time of transfection in (E).