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. 2023 Mar 22;8(6):e162436. doi: 10.1172/jci.insight.162436

Figure 5. YTHDF2 recognizes specific m6A modification sites in the MCP1 CDS region.

Figure 5

(A) Schematic representation of m6A sites of MCP1 mRNA in the CDS region. (B) Schematic representation of mutated m6A sites of MCP1 mRNA in the pCDNA3.1 vector. Adenines in chr17:34256757 (MCP1-Mut1) and chr17:34256822 (MCP1-Mut2) were mutated to guanine. (C) pCDNA3.1 vectors expressing the MCP1 mutant were transfected into 293T cells treated with siNC or siYTHDF2, and the relative MCP1 mRNA expression was quantified by qPCR (n = 9). (D) Representative blot images of MCP1 in siNC- or siYTHDF2-treated 293T cells transfected with MCP1-mutant pCDNA3.1 vectors (n = 9). (E and F) YTHDF2 RIP-qPCR analysis of MCP1 mRNA in the MSCs (n = 9) transfected with MCP1-mutant pCDNA3.1 vectors. Data are presented as the mean ± SD. Two-tailed Student’s t test was performed in panels C and E and 1-way ANOVA followed by Bonferroni’s test was performed in F. *P < 0.05; ****P < 0.0001. NS, not significant; MSCs, mesenchymal stem cells; MCP1, monocyte chemoattractant protein 1; siNC, control siRNA; siYTHDF2, siRNA for YTHDF2.