FIG 3.

DTMUV NS2A inhibits the virus-induced IFN signaling pathway. (A, B) DEFs were transiently transfected with each of DTMUV-derived expression plasmids (400 ng/well) and subsequently transfected with pRL-TK plasmid (40 ng/well), pGL3-IFN-β-Luc (400 ng/well), or pGL4-ISRE-Luc (400 ng/well). At 24 h posttransfection, cells were infected with DTMUV (25 μl containing 100 TCID₅₀ per well), and the IFN-β/ISRE-Luc activity was measured at 36 h postinfection. (C, D) Dose-dependent analysis of the inhibition of DTMUV-mediated IFN-β/ISRE promoter activity by both pNS1 and NS2A. DEFs were transiently transfected with the indicated amount of pNS1 and NS2A plasmid (100 ng, 200 ng, or 400 ng/well) and subsequently transfected with pRL-TK plasmid (40 ng/well), pGL3-IFN-β-Luc (400 ng/well), or pGL4-ISRE-Luc (400 ng/well). At 24 h posttransfection, cells were infected with DTMUV (25 μl containing 100 TCID₅₀ per well); the IFN-β/ISRE-Luc activity was measured at 36 h postinfection. All data are represented as the mean ± SEM (n = 4). Significant differences from the mock groups are indicated by *, P < 0.05; **, P < 0.01; ***, P < 0.001.