Fig. 6: Aberrant activation of TGF-β1-Smads signaling in lens of high myopia.

a Detection of TGF-β1 and growth hormone levels in human lens epithelium by growth factor array (n = 5, both p = 0.003). b Examination of protein expression by Western blotting of TGF-β1, TGF-βR1, and downstream Smad2/3, p-Smad2/3, and Smad4 (n = 3, p = 0.012, p = 0.013, p = 0.019, p = 0.004, and p = 0.019, respectively). Right, the band density in Western blotting was normalized to loading control as a ratio for statistical analysis. c Immunofluorescence images of TGF-βR1 staining in human lens epithelium (n = 5). Scale bar: 50 μm. d Measurement of human TGF-β1 concentration with ELISA in aqueous humor (n = 28 in the control group vs. n = 31 in the highly myopic group, p = 0.876) and primary lens epithelial cell culture supernatant (3, 5, and 5 different cultures in the blank, emmetropic control, and highly myopic group, respectively, one piece of lens epithelium in one culture for this assay, bland vs. emmetropia p = 0.164, emmetropia vs. high myopia p = 0.027, blank vs. high myopia p = 0.002). Blank refers to DMEM without culture of lens epithelium. e Western blotting of protein expression of TGF-β1, TGF-βR1, Smad2/3, and Smad4 (n = 3, p = 0.012, p = 0.041, p = 0.048 and p = 0.008, respectively) and ELISA test of TGF-β1 level in mouse lens epithelium of defocus-induced highly myopic eye and the contralateral eye (n = 6, p = 0.016). Middle, the band density in Western blotting was normalized to loading control as a ratio for statistical analysis. f Western blotting of protein expression of TGF-β1, TGF-βR1, Smad2/3, and Smad4 (n = 3, p = 0.021, p = 0.040, p = 0.032, and p = 0.033, respectively) and ELISA test of TGF-β1 level in lens epithelium of Irbp KO mice and wild type C57BL/6J (n = 3, p = 0.015). Middle, the band density in Western blotting was normalized to loading control as a ratio for statistical analysis. n = biological replicates. In (a, b, e, and f), pooled samples were used. Results are expressed as mean ± SD. Level of significance was detected using two-sided Student’s t test (Figs. a, b, and f) and two-sided paired t test (Fig. e). In (d), TGF-β1 concentration in aqueous humor was analyzed using two-sided Student’s t test, while the concentration in culture supernatant was analyzed using one-way ANOVA with Turkey’s multiple comparisons test for further comparison between two groups. **p < 0.01, *p < 0.05 and ns represents no significant difference. Source data are provided as a Source Data file.