FIG 5.

DEV UL41 inhibits the cGAS-STING pathway by downregulating IRF7 expression. (A) DEFs were cotransfected with IFN-β-luc reporter, pRL-TK, and cGAS-HA, STING-HA, TBK1-HA, or IRF7-HA along with empty vector or UL41-Flag expression plasmid as indicated. Cells were harvested 36 h after transfection and subjected to dual-luciferase reporter assays. All controls and treated groups were performed and examined in triplicate. (B) 293T cells were cotransfected with IRF7-HA and UL41-Flag plasmids. Twenty-four hours after transfection, cells were harvested and subjected to Western blot analysis. (C) DEFs were transfected with UL41-Flag, UL7-Flag plasmid, or the empty vector. Twenty-four hours after transfection, cells were harvested and subjected to Western blot analysis with the indicated antibodies. (D) DEFs were infected with DEV-WT or DEV-dUL41 (MOI = 0.01). Twenty-four hours after infection, cells were harvested and analyzed with qRT-PCR. (E) DEFs were infected with DEV-WT or DEV-dUL41 (MOI = 0.01). Twenty-four hours after infection, cells were harvested and subjected to Western blot analysis with the indicated antibodies. (F) 293T cells were cotransfected with IRF7-HA and UL41-Flag plasmids. Twenty-four hours after transfection, cells were treated with dimethyl sulfoxide (DMSO), MG132 (10 μM), NH₄Cl (20 mM), and 3-MA (10 mM) for 12 h. Protein expression was measured using Western blotting. (G) Vero cells were cotransfected with IRF7-HA and UL41-Flag plasmids, and cells were subjected to Western blot analysis 36 h after transfection. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ns, no significant difference.