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. 2021 May 27;8(15):2100606. doi: 10.1002/advs.202100606

Figure 1.

Figure 1

METTL14‐mediated m6A modification inhibits RLR‐induced innate immunity signaling. A) Immunoblot analysis of METTL14 in Mettl14 +/+ and Mettl14 +/− peritoneal macrophages. B) m6A dot blot assays of Mettl14 +/+ and Mettl14 +/− peritoneal macrophages, methylene blue staining (as loading control). C) qPCR analysis of Ifnb1, Il6, or Ifna4 mRNA in Mettl14 +/+ and Mettl14 +/− peritoneal macrophages, followed by infection with SeV or EMCV or stimulation with 5′‐ppp RNA for the indicated times. D) Immunoblot analysis of phosphorylated and total IRF3 and TBK1 in lysates of Mettl14 +/+ and Mettl14 +/− peritoneal macrophages infected with SeV for the indicated times. E) Immunoblot analysis of phosphorylated and total IRF3 and TBK1 in lysates of Mettl14 +/+ and Mettl14 +/− peritoneal macrophages infected with EMCV for the indicated times. F) Immunoblot analysis of METTL14 in Mettl14 +/+ and Mettl14 +/− BMDMs. G) m6A dot blot assays of Mettl14 +/+ and Mettl14 +/− BMDMs, methylene blue staining (as loading control). H) qPCR analysis of Ifnb1, Il6, or Ifna4 mRNA in Mettl14 +/+ and Mettl14 +/− BMDMs, followed by infection with SeV or stimulation with 5′‐pppRNA for the indicated times. I) Immunoblot analysis of phosphorylated and total IRF3 and TBK1 in lysates of HEK293T cells transfected with control plasmid or plasmid expressing Flag‐METTL14 (above blots) for 24 h, followed by infection with SeV for the indicated times. J) Immunoblot analysis of phosphorylated and total IRF3 and TBK1 in lysates of HEK293T cells transfected with control plasmid or plasmid expressing Flag‐METTL14 or Flag‐METTL14 (R298P) (above blots), followed by SeV infection for 8 h. Data information: Data are presented as mean ± S.D. Two‐tailed unpaired Student's t‐test; *P < 0.05; **P < 0.01 (C,H).