Fig 1: Hepatic BMP8B mRNA expression in mice.Liver tissues were collected from 10 weeks mice fed with chow and CDHF diet. Further, BMP8B mRNA expression was measured by using RT-PCR. Fold change was normalized to 18S rRNA. Data were expressed as mean ± SD. *p<0.05 was considered statistically significant with the corresponding group. A) WT (n = 5) vs WT + CDHF (n = 5). B) iNOS-/- (n = 5) vs iNOS-/- + CDHF (n = 5). Abbreviations: BMP8B: Bone morphogenetic protein-8B; CDHF: Choline deficient high fat; WT = wild type; iNOS-/-: inducible nitric oxide synthase knock out.
Fig 2: Serum concentration of BMP8B.BMP8B concentration (pg/ml) was measured in serum samples of control (n = 56), NAFL (n = 72), and NASH (n = 77) patients by using an ELISA kit. Kruskal-Wallis One-way ANOVA followed by Dunn’s multiple comparison test was applied to check the statistical significance between the groups. ***p = 0.0001, #p < 0.0001. Abbreviations: BMP8B: Bone morphogenetic protein-8B; NAFLD: Non-alcoholic fatty liver disease; NASH: Non-alcoholic steatohepatitis; ELISA: enzyme-linked immunosorbent assay.
Fig 3: Immunohistochemistry (IHC) analysis of BMP8b expression.IHC analysis was performed in liver tissues of healthy controls and NASH patients. Liver tissues were stained with Hematoxylin and eosin (H&E) staining and then immunostained with BMP8b antibody. A) Control (n = 4), B) F0—No fibrosis (n = 4), C) F2—Moderate fibrosis (n = 4), D) F3—Severe fibrosis (n = 4). The arrows indicate the immune reaction in hepatocytes.
Fig 4: Hepatic BMP8b mRNA expression.RT-PCR was used to measure BMP8b mRNA expression in liver tissues of control subjects (n = 4) and NASH patients (n = 5). Fold change was normalized to 18S rRNA. Data were expressed as mean ± SD. *p < 0.05 was considered statistically significant. Abbreviations: BMP8B: Bone morphogenetic protein-8B; NASH: Non-alcoholic steatohepatitis; RT-PCR: Real-time polymerase chain reaction.
Fig 5: Receiver operator characteristic (ROC) curve analysis of BMP8B.Serum BMP8B concentrations of healthy controls and NASH patients were used for ROC curve analysis. AUC = 0.96; sensitivity = 92.21%; specificity = 92.73%.
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