FIGURE 2.

Effect of HCV defective genomes on viral replication and release. (A) Effect of IFDMs on HCV replication. The pFK-I₃₄₁PI-Luc/NS3-3′/JFH1–replicating construct (replicon wt) was electroporated in Huh7.5 cells simultaneously with different RNA IFDMs constructs at a ratio 1:1. pHPI 8388 clone (containing the FL HCV sequence) served as a control. The majority of the IFDMs used were HCV genotype 3a (8381, 8387, 8389) and 1b (32A). Cells were collected at 48 h, lyzed and luciferase activity was measured and normalized to total protein. The value for replicon wt was arbitrarily set as 100% and all other values are a percentage of this (seven independent experiments). (B) Quantity of viral RNA (viral copies/mL) from replicon wt, control (containing the FL HCV sequence) and IFDMs 8381, 8387, 8389, 32A measured in the cell culture supernatants of the above experiment with the SACACE Real Time PCR kit for HCV detection. (C) HCV Ag (Core) was measured in cell lysates and supernatants from replicon wt, control (containing the FL HCV sequence) and IFDM HCV 3a (8389) using the Abbott ARCHITECT HCV Ag assay. ^∗p-value ≤ 0.05; ^∗∗p-value ≤ 0.005; ^∗∗∗p-value ≤ 0.001.