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. 2020 Dec 4;10:21280. doi: 10.1038/s41598-020-77978-z

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© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Effects of orally administered BR103354 in CDAHFD mice. C57BL/6J mice were fed normal chow diet (n = 5) or choline-deficient, L-amino acid-defined, high-fat diet (CDAHFD) for 4 weeks to induce NASH (n = 8–10/group). The vehicle, BR103354 (10 and 30 mg/kg) and LJN (positive control, 3 mg/kg) were orally administered for 10 weeks. (a) Relative FAP activity was assessed by FRET-quench and serum FGF21 level at week 14. (b) The plasma levels of ALT (GPT), AST (GOT), total cholesterol, glucose and TG in NCD and CDAHFD mice were measured at week 14. (c) Steatosis scores ranged from 0 to 3 (normal = 0, minimal = 1, moderate = 2, marked = 3) were determined. Hepatic steatosis and fibrosis (Picrosirius red stain) were analyzed in a blinded manner or by digital image analyzer. (d–e) Histological evaluations of liver tissue were performed in each group. Paraffin-embedded liver sections were stained with H&E (d) and Sirius red (e). Representative photographs of each group were presented. Images were obtained at 100x (Scale bar = 50 µm). All data are the means ± S.D. *P < 0.05, **P < 0.01 and ***P < 0.001 vs. CDAHFD-vehicle group and †P < 0.001 vs. NCD-vehicle group.

Effects of orally administered BR103354 in CDAHFD mice. C57BL/6J mice were fed normal chow diet (n = 5) or choline-deficient, L-amino acid-defined, high-fat diet (CDAHFD) for 4 weeks to induce NASH (n = 8–10/group). The vehicle, BR103354 (10 and 30 mg/kg) and LJN (positive control, 3 mg/kg) were orally administered for 10 weeks. (a) Relative FAP activity was assessed by FRET-quench and serum FGF21 level at week 14. (b) The plasma levels of ALT (GPT), AST (GOT), total cholesterol, glucose and TG in NCD and CDAHFD mice were measured at week 14. (c) Steatosis scores ranged from 0 to 3 (normal = 0, minimal = 1, moderate = 2, marked = 3) were determined. Hepatic steatosis and fibrosis (Picrosirius red stain) were analyzed in a blinded manner or by digital image analyzer. (d–e) Histological evaluations of liver tissue were performed in each group. Paraffin-embedded liver sections were stained with H&E (d) and Sirius red (e). Representative photographs of each group were presented. Images were obtained at 100x (Scale bar = 50 µm). All data are the means ± S.D. *P < 0.05, **P < 0.01 and ***P < 0.001 vs. CDAHFD-vehicle group and †P < 0.001 vs. NCD-vehicle group.