Fig. 3

BMP9 expression is regulated by C/EBPα in vitro. A IF staining for BMP9 and F4/80 in liver slices from young and aged mice treated with PBS or APAP (300 mg/kg) for 24 h. B, C Primary hepatocytes and MΦs were isolated from young and aged mice by in situ perfusion of mouse livers. Protein expression levels of BMP9, C/EBPα, ATG3 and ATG7 in mouse primary hepatocytes (B) and hepatic MΦs (C). D–G To study the regulation of BMP9 by C/EBPα in MΦs, we altered Cebpa expression in iBMDMs. D Protein expression levels of C/EBPα and BMP9 after overexpressing Cebpa. E Protein expression levels of C/EBPα and BMP9 after inhibiting Cebpa with or without treatment with the APAP-treated AML-12 cell supernatant (Sup). F Protein expression levels of CD206 and iNOS after overexpressing Cebpa with or without Sup treatment. G mRNA expression of related cytokines (Il1b, Il6 and Tnfa) after overexpressing Cebpa with or without Sup treatment. (H, I) To study the regulation of BMP9 by C/EBPα in hepatocytes, we altered Cebpa expression in AML-12 cells. H Protein expression levels of C/EBPα and BMP9 after overexpressing Cebpa. I Protein expression levels of C/EBPα and BMP9 after inhibiting Cebpa with or without Sup treatment. The average target gene/Gapdh ratios in different experimental groups relative to the control group. *p < 0.05, **p < 0.01, and ***p < 0.001