Fig. 2: TGF-β1 correlates with HMC severity and induces increased m6A modifications in an METTL3-dependent manner.

a The dot blot assay of m6A abundance (n = 3, p = 0.0130). b Colorimetric quantification of relative m6A abundance (n = 12, p = 0.0022). c Gene expressions of m6a writers (n = 12, METTL3 p < 0.0001, WTAP p > 0.9999, METTL14 p = 0.5750, ALKBH5 p > 0.9999, and FTO p > 0.9999). d METTL3 protein expression in ARC and HMC (n = 3, p = 0.0352). e Nuclear staining of METTL3 in ARC and HMC (n = 3, p = 0.0290). The nuclear counter stain is DAPI. Scale bar: 50 μm. f METTL3 gene expression in cells treated with vehicle, 100 μM or 200 μM H₂O₂ (n = 3, 100 μM p = 0.0945, 200 μM p = 0.0570). g METTL3 gene expression in cells treated with vehicle or 10 ng mL⁻¹ TGF-β1. (n = 3, p = 0.0345) h METTL3 protein expression in cells treated with vehicle or 10 ng mL^-1 TGF-β1 (n = 3, p = 0.0297). i, j The dot blot assay of m6A abundance in cells with defferent treatments. (n = 3, i: NC + TGF-β1 p = 0.0072, METTL3 OE + TGF-β1, p = 0.0003; j: NC + TGF-β1 p = 0.0360, METTL3 OE + TGF-β1 p = 0.0456). k Preoperative anterior segment OCT images of lenses from ARC and HMC patients. Orange and yellow dotted circles indicate the cortex and nucleus, respectively. Average nuclear intensity was noted at bottom right. Correlation between preoperative nuclear intensity and their LEC TGF-β1 concentrations is shown. (n = 9, p = 0.0318). The band or dot density was normalized to loading control as a ratio for statistical analysis. Data present are mean ± SD. Level of significance was detected using two-sided Student’s t test (a, b, d, e, g, h), one-way ANOVA with Dunnett’s multiple comparisons test (f, i, j), two-way ANOVA with Šídák’s multiple comparisons test (c), and Pearson’s correlation analysis (k). n = biological replicates, ns. = not significant, ****p < 0.0001, ***p < 0.001, **p < 0.01 and *p < 0.05.