FIGURE 2.

IRF3 deficiency abrogates viral induction of ZAP. A, left panel, HeLa and HeLa cells stably expressing BVDV Npro (HeLaNpro) were mock infected or infected with 100 HAU/ml of SeV for 16 h. The cell lysates were immunoblotted for Npro (using anti-HA monoclonal antibody), IRF3, actin, and ISG56. Right panel, HeLa and HeLaNpro cells were mock treated or treated with 100 nm of epoxomicin for 12 h prior to immunoblot analysis of Npro, IRF3, and actin. B, Q-PCR analysis of ZAP, IFN-β, and OAS1 mRNA levels in HeLa and HeLaNpro cells mock treated or stimulated with 500 units/ml IFNα, 100 HAU/ml of SeV or transfected with 2 μg of in vitro transcribed HCV RNA for 8 h. mRNA abundance was normalized to cellular 28 S ribosomal RNA. Fold changes were calculated by dividing normalized mRNA abundance following various treatments by that of the mock treated HeLa cells.