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. 2024 Sep 30;13(19):e034470. doi: 10.1161/JAHA.124.034470

Figure 6. IGF2BP2 enhances PTEN mRNA expression through a m6A‐dependent pathway.

Figure 6

A, B, The protein and mRNA expression of PTEN in PASMCs after the siRNA‐induced silencing of IGF2BPs, as determined by qRT‐PCR and Western blot (nitrocellulose, n=4–6; si‐IGF2BPs, n=4–6). C, RIP analysis with an IGF2BP2‐specific antibody to detect the enrichment of IGF2BP2 binding to PTEN mRNA (n=4). D, An RNA pull‐down assay validated the direct interaction between IGF2BP2 and PTEN mRNA in PASMCs. E, The expression of IGF2BP2 was positively correlated with the PTEN expression in hypoxic mice. F, G, In PASMCs, METTL3 overexpression enhanced the PTEN protein and mRNA expression, whereas IGF2BP2 siRNA reversed the upregulation of PTEN protein and mRNA expression, as determined by western blot and qRT‐PCR (Nor, n=6–7; HYP, n=6–7; HYP+METTL3, n=6–7; HYP+METTL3+si‐IGF2BP2, n=6–7; HYP+METTL3+si‐nitrocellulose, n=6–7). Each data point in the figure represents a unique biological replicate. Statistical analysis was performed with 1‐way ANOVA. The data are presented as the mean±SD. *P<0.05, **P<0.01, ***P<0.001. HYP, hypoxia; IGF2BP, insulin‐like growth factor 2 mRNA‐binding protein; METTL, methyltransferase‐like; Nor, normal; PASMC, pulmonary artery smooth muscle cell; PI, propidium iodide; PTEN, phosphate and tension homology deleted on chromosome 10; qRT‐PCR, quantitative real‐time polymerase chain reaction; RIP, RNA immunoprecipitation; and siRNA, small interfering RNA.