Fig. 7

Glutaminolysis regulated ESCs activation via activated mTORC1 pathway. A, B Western blot (A) and quantification analysis (B) of p-p70-S6K, p-S6, p-4E-BP1 protein level in ESCs after pretreatment with BPTES for 6 h and TGF-β1 stimulation for another 48 h. C, D Western blot (C) and quantification analysis (D) of p-p70-S6K, p-S6, p-4E-BP1 protein level in ESCs treated with TGFβ1 in the presence or absence of glutamine for 48 h. E Quantitative PCR analysis of αSMA and Collagen I mRNA levels in ESCs after pretreatment with gradient doses of Rapamycin (Rapa, mTOR inhibitor) or 0.1% DMSO for 12 h and TGF-β1 stimulation for another 48 h. F, G Western blot (F) and quantification analysis (G) of α-SMA and Collagen I protein levels in ESCs pretreated with 200 nM Rapamycin or 0.1% DMSO for 12 h and treated with TGF-β1 stimulation for another 48 h. 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). F The OCR analysis of ESCs pretreated with vehicle (0.1% DMSO) or Rapa (200 nM) for 12 h and then treated together with 10 ng/ml TGF-β1 for 48 h (n = 5 in each group). Statistical analyses were performed by Student’s t-test. *p < 0.05, **p < 0.01, ***p < 0.001