Fig. 6

Analysis of mitochondrial function in glutaminolysis inhibition experiments. A Relative MMP analysis of ESCs treated with vehicle (0.1% DMSO) or BPTES (10 µM) for 6 h and then treated together with or without 10 ng/ml TGF-β1 for 48 h. B Relative MMP analysis of ESCs treated with or without 10 ng/ml TGF-β1 in the presence or absence of 2 mM glutamine for 48 h. C Relative MitoSOX fluorescence intensity of ESCs treated with or without 10 µM BPTES and 10 ng/ml TGF-β1. D Relative MitoSOX fluorescence intensity of ESCs treated with or without 2 mM glutamine and 10 ng/ml TGF-β1. E, F Representative images of MitoSOX staining in ESCs from each group. The scale bar = 150 μm. Data represent at least 3 biologically independent experiments. Treatment groups compared with their control (set at 1) was analyzed using a one-sample Student’s t-test, and subsequent comparisons of treatment groups was performed using the ANOVA followed by Turkey post-hoc analysis. G Relative mitochondrial ATP levels of ESCs treated with or without 10 µM BPTES and 10 ng/ml TGF-β1. H The OCR analysis of ESCs treated with or without 10 µM BPTES and 10 ng/ml TGF-β1. I Relative mitochondrial ATP levels of ESCs treated with or without 2 mM glutamine and 10 ng/ml TGF-β1. J The OCR analysis of ESCs cultured in Gln-containing or Gln-free medium for 48 h. To measure the acute response of ESCs to glutamine, ESCs were cultured in Gln-free medium, and glutamine was added back when OCR was measured. Data represent at least 3 biologically independent experiments. Statistical analyses were performed by one-way ANOVA plus Tukey’s multiple comparisons test. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001