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. 2016 Apr 27;291(25):13206–13215. doi: 10.1074/jbc.M116.718080

FIGURE 5.

FIGURE 5.

IRF7 binds to USP25 promoter. A, a schematic model of USP25 promoter and its mutations. B, HEK293 cells were transfected with the indicated luciferase (Luc.) reporter (100 ng) together with Vec, IRF7 (0.1 μg), IRF3 (0.5 μg) (left graph), p65, p50, or p52-RelB (0.2 μg) (middle graph). Twenty hours later, luciferase reporter assays were performed. HEK293 cells were transfected with the indicated luciferase reporter (100 ng). Twenty hours later, cells were infected with SeV for 8 h followed by luciferase reporter assays (right graph). C, HEK293 cells were transfected with the indicated luciferase reporter (100 ng) together with Vec, IRF7 (0.02–0.1 μg), or IRF3 (0.1–0.5 μg) (left graph). Twenty hours later, luciferase reporter assays were performed. Immunoblotting analysis was performed to examine the expression of transfected plasmids (right panels). D, Irf3−/−Irf7−/− MEFs were reconstituted with the empty vector (Irf3−/−Irf7−/− + Vector), IRF3 (Irf3−/−Irf7−/− + FLAG-IRF3), or IRF7 (Irf3−/−Irf7−/− + FLAG-IRF7) through lentivirus-mediated gene transfer. Cells were left untreated or infected with SeV for 12 h followed by ChIP analysis. TSS, transcription start site. Data shown are representatives of four (B) or three independent experiments (C and D). *, p < 0.05; **, p < 0.01; ***, p < 0.001. Error bars represent S.D. Rel., relative.