Fig. 3

Inhibition of glutaminolysis suppressed α-SMA and Collagen I expression in TGF-β1–induced ESCs. A–C ESCs were 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 (A). The changes in α-SMA and Collagen I mRNA levels were determined by quantitative PCR analysis (B), changes in α-SMA and Collagen I protein levels were determined by Western Blot analysis (C). D–F ESCs were treated with or without 10 ng/ml TGF-β1 in the presence or absence of 2 mM glutamine for 48 h (D). The changes in mRNA levels (E) and protein levels (F) of α-SMA and Collagen I were determined. G-I ESCs were treated with or without 10 ng/ml TGF-β1 for 48 h and switched to Gln-free or Gln-containing medium for 24 h after TGF-β1 treatment (G). The changes in mRNA levels (H) and protein levels (I) of α-SMA and Collagen I were determined. J–L ESCs were treated with 10 ng/ml TGF-β1 in the presence or absence of 2 mM Gln or 2 mM α-KG for 48 h (J). The changes in mRNA levels (K) and protein levels (L) were determined. 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 parametric analysis (student’s t test or ANOVA followed by Turkey post-hoc analysis). *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. n = 4 in each group