Setdb2 suppresses the expression of antiviral genes and cytokine production nu BM-Mϕ.

Gene expression (A, C) and cytokine production (B) by control and Setdb2^-/- BM-Mϕ stimulated with IFN-I for 24 hours. Data represents the mean ± SEM of 3 independent experiments. (A) Comparison of antiviral and proinflammatory genes by RT-PCR. Values on scatter plot represent log₁₀ (Δ^Ct). The gray lines represent a 2-fold change in gene expression; upregulated genes (red dots), downregulated genes (green dots). (B) Concentration of IL-1β, IL-6, IL-10, IL-12p40, G-GSF, and TNF-α in the supernatants of control and Setdb2^-/- BM-Mϕ. (C) RT-PCR of Isg15 and Mx1. Data were normalized to control BM-Mϕ. (D-G) Control and Setdb2^-/- BM-Mϕ were treated with a vehicle control, IFN-I (D-G), or IFN-II (F, G) for 24 hours. Data are mean ± SEM relative to IgG control; n = 2–3 experiments (D, E) ChIP analysis of Setdb2 binding and H3K9 tri-methylation in the Isg15(D) and Mx1(E) promoter. (F, G) ChIP analysis of STAT1 (F) and IRF7 (G) binding in the Mx1 promoter. (H) Control and Setdb2^ffLyz2^cre+ mice inoculated with a sublethal dose of IAV (1 x 10⁴ PFU).) RT-PCR of Rnasel, Pkr, Mx1, and Isg15 in infected lungs on day 4 post-infection. Data are mean ± SEM (n = 6–14 mice) from 2–3 independent experiments. *p<0.05; **p<0.01; ***p<0.001.