Measurement of HCV replication dynamics.

<p>(A) Schematic representation of the subgenomic HCV luciferase reporter replicon used for the study. The 5′-non-translated region (NTR) contains the HCV internal ribosome entry site (IRES), controlling translation of the firefly luciferase gene (Luc). The non-structural proteins of the HCV isolate JFH1 are under control of the encephalomyocarditis (EMCV) virus IRES, and are followed by the orthologous 3′- NTR of JFH1. (B&C) Quantitative assessment of the HCV replication dynamics upon instantaneous (t = 0 h) electro-transfection into (B) high permissive Huh7-Lunet cells or (C) low permissive Huh-7 low passage cells. The top panel shows a Northern blot analysis of the viral plus- and minus-strand RNA. The lower panel shows a graph of the Northern blot signals quantified by phosphor imaging (plus-strand RNA: blue lines; minus-strand RNA: red lines), as well as the corresponding luciferase activity (RLU, yellow lines). Luciferase activity and plus-strand RNA are normalized to the input values (2 h and 0 h, respectively; one representative experiment is shown. Lines in the plots are for illustrative purposes and connect data points, but are not results of mathematical modeling.