Figure 6

Preparation of stable replicon cell line replicating HCV 2a sub-genomic RNA. Interferon sensitive (S3-GFP) and interferon resistant (R4-GFP) Huh-7 cells were transfected with pSGR-GFP replicon RNA and then selected with G-418 (500 μg/ml). Single G-418 resistant cell clones were picked and stable cell lines were generated. (A) Intracellular GFP expression in S3-GFP and R4-GFP stable replicon cell lines. (B) Quantification of GFP expression in these IFN-sensitive (S3-GFP) and resistant (R4-GFP) cells was performed flow analysis. Approximately 81% of S3-GFP and 84% of R4-GFP cells in the culture showed intracellular GFP expression. High-level expression of HCV-GFP was seen in both cell lines suggesting that both the sensitive and resistant Huh-7 clones support high level HCV replication.