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. 2012 Aug 6;7(8):e42609. doi: 10.1371/journal.pone.0042609

Figure 1. The gt 1a/2a-Rluc virus is capable of multi-cycle virus growth amenable to unbiased inhibitor detection.

Figure 1

A. Schematic of the genotype 1a/2a-Rluc virus which harbors Core-NS2 from the genotype 1a H77 isolate fused to NS3-NS5B of the genotype 2a JFH1 isolate with an Rluc reporter gene cloned between NS5A and NS5B. B–E. Huh-7.5 cells were infected with gt 1a/2a-Rluc (MOI = 0.05) and processed for HCV Core immunofluorescence or Renilla Luciferase expression 48, 72 and 96 h post-infection (pi). Virus spread was inhibited using an HCV entry inhibitor (EI; 1 µM) added 16 h post-entry [EI (Post-Entry)] or with an SPP inhibitor (LY411575; 0.5 µM). Virus infectivity and spread were assessed directly using immunofluorescence microscopy of HCV Core (green) and Huh-7.5 nuclei (red) to calculate the number of infected cells per viral foci (B, C, & D) or indirectly with Renilla Luciferase (D & E). Results are expressed as the mean and standard deviation of at least two independent assays.