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. 2021 Feb 22;218(3):e20201798. doi: 10.1084/jem.20201798

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© 2021 Gao et al.

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Figure 7. — Ifnb production is inhibited by single-nucleotide methylation of its promoter. (A) Schematic picture showing the strategy for generating CpG mutant knock-in mice (Ifnb1^CpG(G-A)). (B) The mRNA levels of Ifnb induced by different viruses in WT and Ifnb1^CpG(G-A) BMDMs were measured by qPCR. ChIP assays were performed and quantified by qPCR to detect the binding of IRF3 to the WT or Ifnb1^CpG(G-A) promoters (n = 4). (C) WT and Ifnb1^CpG(G-A) mice (6–8 wk) were i.n. infected with a sublethal dose (0.1 HA) of H5N1 influenza virus (n = 4). (D and E) The body weight loss (D) and survival rate (E) were measured for 14 d (n = 18). (F) Viral titers in the lung were quantified on day 2 using the TCID₅₀ assay (n = 4). (G) ELISA for IFN-β in the sera of WT and Ifnb1^CpG(G-A) mice infected with H5N1 influenza virus on days 2 and 5 (n = 5). (H–K) WT and Uhrf1^MKO mice bred on the Ifnb1^CpG(G-A) background were i.n. infected with a sublethal dose (0.1 HA) of H5N1. The mRNA levels of Ifnb induced by different viruses in the indicated BMDMs were measured by qPCR (H; n = 3). The body weight loss (I; n = 8), survival rate (J; n = 8) for 14 d, and viral titer (K; n = 5) on day 2 are shown. The data in the qPCR assay are presented as fold changes relative to the actin mRNA levels. All data are representative of at least three independent experiments. The data are presented as means ± SEMs. The significance of differences was determined by a t test. *, P < 0.05.