Fig. 1

Replication of TPF1 and sAH RNAs in transfected Huh7 cells. a Structure of the subgenomic replicon. b Huh7 cells were transfected with the wild-type replicons (WT) or a replicon RNA carrying the adaptive mutations (Recovered). Numbers below the plates refer to colony-forming units (CFU) per microgram of in vitro-transcribed replicon RNA. With the rep-TPF1 of WT replicons, ten independent transfections were performed. With the rep-sAH of WT and adapted replicons, three independent transfections were performed. c The structures of the full-length HCV genomes TPF1-4B and sAH-N3.5 are shown. Both full-length genomes carry the heterologous sequence (shaded area) derived from the highly adapted subgenomic replicons, showing the location of the adaptive mutations. GDD is the active-site motif of NS5B polymerase. d–f Time course of HCV RNA replication (d), core Ag expression (e), and core Ag secretion (f) in transfected Huh7 cells. Huh7 cells were electroporated with TPF1-4B (□), sAH-N3.5 (△), and TPF1-ΔGDD (○) RNA transcripts. TPF1-ΔGDD contains a deletion of the polymerase active-site motif (GDD) in NS5B polymerase. The HCV RNA and core Ag levels were measured by qRT-PCR and sandwich EIA at the indicated time points, respectively. Results represent the mean of three independent experiments ± standard deviations