Fig. 6.

Adropin increased CBP expression and its binding with Nrf2 to enhance Nrf2 transcriptional activity. Primary murine hepatocytes pretreated with PA (400 µM) were treated with adropin at different dosages (0–100 ng/ml) for 24 h. The mRNA expression of Cbp (A) and the protein expression of CBP (B-C) were measured. CBP-Nrf2 and CBP-NFκB interactions were studied by immunoprecipitation (IP) in the hepatocytes treated with or without adropin (100 ng/ml) (D-G). Chromatin immunoprecipitation (ChIP) assays were performed to investigate the presence of CBP at the antioxidant response element (ARE) sites on the promoters of Gclc (H) and Gpx1 (I) in the hepatocytes treated with or without adropin (100 ng/ml). Primary murine hepatocytes pretreated with PA (400 µM) were treated with or without adropin (100 ng/ml) and transfected with CBP siRNA or control for 24 h. And the DNA-binding activity of Nrf2 (J) and the mRNA expression of Gpx1 (K) were measured. (A, C, E, F, G) PA-treated group was set as 1. (H, I, J, K) Blank control group was set as 1. The data are expressed as the mean ± SD, n = 3–5, *P < 0.05 versus PA-treatment group. NC indicates negative control. N.S indicates no significance. Adropin-KO mice and the wild type (WT) littermate were fed with MCD or WD for 4 or 16 weeks. The mRNA (L) and protein expression (M-O) of Nrf2 and Cbp were measured. And the liver lysates DNA-binding activity of Nrf2 was detected (P). (L, N, O, P) WT control group was set as 1. The data are expressed as the mean ± SD, n = 6, *P < 0.05 versus WT control.