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. 2020 Apr 29;9(5):1101. doi: 10.3390/cells9051101

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© 2020 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

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Effects of EX-527 on the expression of p-Akt and p-PTEN in the liver of HFD-fed rats. (A) Protein expression of tensin homolog (PTEN), p-PTEN, protein kinase B (Akt), and p-Akt was measured by Western blot analysis using an experimental model of HFD-induced hepatic fibrosis. β-Actin expression was used as the loading control. (B) The intensity of the bands was analyzed densitometrically by ImageJ software. Effect of EX-527 on TGF-β/Smads-related proteins in the liver of HFD-induced diabetic rats by Western blotting analysis. (C) Expression of TGF-β1, Smad2/3, p-Smad2/3, and Smad4 was measured by Western blot analysis in the liver tissue of HFD-induced diabetic rats. β-Actin expression was used as the loading control. (D) The intensity of the bands was analyzed densitometrically by ImageJ software. (E) Representative Immunohistochemical staining of TGF-β1 in the liver of HFD-fed diabetic rats. Black arrows represent TGF-β1 expression. Original magnification: 200×, scale bar: 100 μm. The Western blot results are representative of three separate experiments. (F) The positive area of TGF-β1 staining in the liver (%). (G) ELISA results of TGF-β concentration in the serum of HFD-induced diabetic rats. Statistical analysis was performed using one-way ANOVA followed by Tukey’s HSD post hoc test for multiple comparisons. * p < 0.05, ** p < 0.01, and *** p < 0.001 compared with the normal diet group (ND). ^## p < 0.01, and ^### p < 0.001 compared with the HFD group.

Effects of EX-527 on the expression of p-Akt and p-PTEN in the liver of HFD-fed rats. (A) Protein expression of tensin homolog (PTEN), p-PTEN, protein kinase B (Akt), and p-Akt was measured by Western blot analysis using an experimental model of HFD-induced hepatic fibrosis. β-Actin expression was used as the loading control. (B) The intensity of the bands was analyzed densitometrically by ImageJ software. Effect of EX-527 on TGF-β/Smads-related proteins in the liver of HFD-induced diabetic rats by Western blotting analysis. (C) Expression of TGF-β1, Smad2/3, p-Smad2/3, and Smad4 was measured by Western blot analysis in the liver tissue of HFD-induced diabetic rats. β-Actin expression was used as the loading control. (D) The intensity of the bands was analyzed densitometrically by ImageJ software. (E) Representative Immunohistochemical staining of TGF-β1 in the liver of HFD-fed diabetic rats. Black arrows represent TGF-β1 expression. Original magnification: 200×, scale bar: 100 μm. The Western blot results are representative of three separate experiments. (F) The positive area of TGF-β1 staining in the liver (%). (G) ELISA results of TGF-β concentration in the serum of HFD-induced diabetic rats. Statistical analysis was performed using one-way ANOVA followed by Tukey’s HSD post hoc test for multiple comparisons. * p < 0.05, ** p < 0.01, and *** p < 0.001 compared with the normal diet group (ND). ^## p < 0.01, and ^### p < 0.001 compared with the HFD group.