Fig. 5

Mitochondria ultrastructure and the expression of proteins related to mitochondrial dynamics in TGF-β1-treated ESCs. A, B MitoTracker staining in ESCs treated without (A) or with (B) TGF-β1 (Scale Bars = 20 μm). C, D MitoTracker staining in ESCs treated without (C) or with (D) TGF-β1 (Scale Bars = 10 μm). E Quantitative analysis of mitochondrial mean branch length using the MINA toolset. 34 TGF-β1-untreated cells (NC) and 31 TGF-β1-treated cells from n = 3 independent experiments were blindly scored. Statistical analyses were performed by Student’s t-test. ****p < 0.0001. F, G TEM images of ESCs without (F) or with (G) TGF-β1 (Scale Bars = 10 μm). The yellow arrows indicate mitochondria. H, I TEM images of ESCs without (H) or with (I) TGF-β1 (Scale Bars = 500 nm). J Quantitative analysis of average mitochondrial major axis length. n = 3; 12–15 mitochondria/cell; 10 cells/per group. Statistical analyses were performed by Mann Whitney test. ****p < 0.0001. K, L Western Blot (K) and quantification analysis (L) of MFN1, MFN2, DRP1, and PINK1 protein levels in ESCs treated without or with TGF-β1 (n = 5 in each group). Statistical analyses were performed by using Student’s t-test. **p < 0.01, ***p < 0.001