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

Figure 5. METTL3 ameliorates PASMC pyroptosis by upregulating PTEN.

Figure 5

A, Cotransfection with PTEN siRNA reversed the effects of METTL3 on PASMCs exposed to hypoxia for 48 h, as indicated by the detection of LDH activity (Nor, n=6; HYP, n=6; HYP+METTL3, n=6; HYP+METTL3+siPTEN, n=6; HYP+METTL3+si‐ nitrocellulose, n=6). B, C, METTL3 ameliorated the upregulation of NLRP3, caspase‐1, interleukin‐1β, interleukin‐18, and ASC at both the protein and mRNA levels induced by hypoxia for 48 h, whereas PTEN siRNA reversed these effects (Nor, n=6; HYP, n=6; HYP+METTL3, n=6; HYP+METTL3+siPTEN, n=6; HYP+METTL3+si‐nitrocellulose, n=6). D, METTL3 overexpression abrogated, whereas PTEN knockdown exacerbated, the abnormal upregulation of positive PI staining in PASMCs upon hypoxia exposure for 48 h. Images of fluorescence staining with PI (red) and Hoechst 33342 (blue). Scale bars=100 μm. Each data point in the figure represents a unique biological replicate. Statistical analysis was performed with one‐way ANOVA. The data are presented as the mean±SD. *P<0.05, **P<0.01, ***P<0.001. ASC indicates apoptosis‐associated speck‐like protein containing a caspase recruitment domain; HYP, hypoxia; METTL, methyltransferase‐like; Nor, normal; PASMC, pulmonary artery smooth muscle cell; PI, propidium iodide; and PTEN, phosphate and tension homology deleted on chromosome 10; siPTEN, small interfering phosphate and tension homology deleted on chromosome 10; siRNA, small interfering RNA.