FIG. 5.

Efficient replication of HCV mutant replicons MT(3A-II) and MT(SL-IV) in the established cell lines. (A) Measurement of HCV RNA levels by real-time PCR. Cells from three established cell lines were harvested, and the total RNAs were purified and used for real-time RT-PCR analyses. The quantity of HCV RNA was normalized with that of GAPDH RNA. (B) Agarose gel electrophoresis of the final PCR products obtained by real-time RT-PCR. The PCR products were analyzed by 2% native agarose gel electrophoresis. Lane No, RT-PCR with no RNA; lanes 1pg and 10pg, RT-PCRs with 1 pg and 10 pg of in vitro-transcribed HCV replicon RNA, respectively; lane M, DNA molecular weight marker; lane 1, naïve Huh-7 cells; lane 2, the parental HCV subgenomic replicon (NK5.1); lane 3, MT(3A-II); and lane 4, MT(SL-IV).