Fig 2: Hepatic Bmp8a expression is upregulated in NAFLD mice. A Representative images of H&E and Sirius Red staining. NAFLD activity score evaluation (upper panel) and quantification of Sirius Red expressed as % of stained area (lower panel). B Hepatic mRNA levels of Col1a1 and Serpin1 determined by RT-qPCR and normalized to 36b4 gene expression. C Hepatic mRNA levels of Bmp8a determined by RT-qPCR and normalized to 36b4 gene expression. D, E, F and G Correlation of matched Bmp8a mRNA expression with NAFLD activity score, fibrosis stage, Col1a1 and Serpin1 mRNA expression, respectively. Experimental conditions: mice fed with HFD or CHD (control group) for 16 weeks (N = 5 animals per group). NAFLD, nonalcoholic fatty liver disease. Data are expressed as fold increase relative to control condition (CHD group, 1) and presented as mean ± SEM. *p < 0.05, **p < 0.01 and ***p < 0.005, HFD vs. CHD

Fig 3: TGFß induces BMP8A expression and secretion in LX2 HSCs and in Huh7 cells. A Representative phase-contrast images. B mRNA levels of COL1A1 and TGFB1 determined by RT-qPCR and normalized to 36B4 gene expression. C Representative blot of cell lysates with aSMA and COL1A1 antibodies. Ponceau staining as loading control. D Representative phase-contrast images. E mRNA levels of BMP8a determined by RT-qPCR and normalized to 36B4 gene expression. F Representative blot of CM with BMP8A antibody. Ponceau staining as loading control. As positive control ( +), HEK293 whole cell lysate was used. Experimental conditions: LX2 or Huh7 cells treated with TGFß 10 ng/ml for 24 h (n = 3 independent experiments performed by duplicate). Huh7, human hepatocyte-derived cell line. LX2, human HSC line. CM, cultured media. Data are expressed as fold increase relative to control condition (C, 1) and presented as mean ± SEM. *p < 0.05, **p < 0.01 and ****p < 0.0001 TGFß vs. C

Fig 4: Increased hepatic Bmp8a expression in BDL mice. A Representative images of H&E and Sirius Red staining. Evaluation of the fibrosis stage (upper panel) and quantification of Sirius Red expressed as % of stained area (lower panel). B Hepatic mRNA levels of Col1a1, Acta2 and Serpin1 determined by RT-qPCR and normalized to 36b4 gene expression. C Hepatic mRNA levels of Bmp8a determined by RT-qPCR and normalized to 36b4 gene expression. D, E, F and G Correlation of matched Bmp8a mRNA expression with fibrosis stage, Col1a1, Acta2 and Serpin1 mRNA expression, respectively. Experimental conditions: mice with either BDL or sham operation and sacrificed 28 days after operation (N = 7 animals per group). BDL, bile duct ligation. Data are expressed as fold increase relative to control condition (sham group, 1) and presented as mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.005 and ****p < 0.0001, BDL vs. Sham

Fig 5: Circulating BMP8A levels are increased in BDL mice. A Serum levels of BMP8A determined by ELISA. Data are expressed as pg/ml and presented as mean ± SEM. B and C Correlation of matched serum BMP8A levels with hepatic Bmp8a mRNA expression and fibrosis stage, respectively. Experimental conditions: mice with either BDL or sham operation and sacrificed 28 days after operation (N = 7 animals per group). BDL, bile duct ligation. **p < 0.01, BDL vs. Sham