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. 2019 Oct 3;158(4):304–313. doi: 10.1111/imm.13116

Figure 4.

Figure 4

FOXO3a directly regulates type IV CIITA transcription. (a) A CIITA type IV promoter luciferase construct (DRA300) was transfected into HEK293 cells with FOXO followed by treatment with interferon‐γ (IFN‐γ). Luciferase activities were normalized by both protein concentration and GFP fluorescence. (b) RAW cells were treated with IFN‐γ and harvested at the indicated time‐points. ChIP assay was performed with anti‐FOXO3a or IgG. (c) RAW cells were treated with IFN‐γ for 12 hr. Re‐ChIP assay was performed with the indicated antibodies. (d, e) RAW cells were transfected with small interfering RNA (siRNA) targeting STAT1 or scrambled siRNA (SCR) followed by treatment with IFN‐γ for 12 hr. Knockdown efficiency was examined by Western blotting. ChIP assay was performed with anti‐FOXO3a. (f–h) RAW cells were transfected with siRNA targeting FOXO3a or SCR followed by treatment with IFN‐γ for 12 hr. Re‐ChIP assay was performed with indicated antibodies. Data represent averages of three independent experiments and error bars represent SEM. (i) A schematic model. *P ≤ 0·05.