FIG 6.

Regulation of HCV-SGR replication in mouse cells by mouse type III IFNs. (A) Phenotypic appearances of all MLT cells. (B) Transient replication of the HCV luciferase replicon without polymerase activity in MLT-MAVS^−/− miR122, MLT-IFNAR^−/− miR122, and MLT-IFNAR^−/− IFNLR1^−/− miR122 cells. At the indicated time points posttransfection, HCV RNA replication was determined by luciferase activity. (C) Indirect immunofluorescence analysis of NS5A expression in given cell lines 48 h posttransfection. The HCV-NS5A protein was detected by specific antibody (9E10), and cell nuclei were stained with DAPI. (D) Effect of exogenously added mouse type I and III IFNs (mIFNa and mIFN-λ3, respectively) on HCV replication in given MLT cells. Forty-eight hours after administration of the given drugs, HCV RNA replication was determined by luciferase assays. Luciferase activity was normalized to that measured in control cells that were mock treated. Asterisks indicate a significant difference compared to each related mock group. (E) Repair of type III IFN receptor expression restores responsiveness of MLT-IFNAR^−/− IFNLR1^−/− miR122 cells to antiviral effect mediated by mIFN-λ3. Given MLT cells carrying a luciferase replicon were transduced to express GFP or the mIFNLR1 cDNA. Subsequently, cells were treated as indicated with 2′CMA or mIFN-λ3. Luciferase activity was determined 48 h later and is expressed relative to that in mock-treated and untransduced cells. Asterisks show a significant difference between the indicated data. All graph data are shown as mean values ± SDs from three independent experiments; images are representative of two individual experiments.