Fig. 5
From: Elevated Kallistatin promotes the occurrence and progression of non-alcoholic fatty liver disease
KAL induces nuclear translocation of NF-κB p65 by reducing its binding to CGI-58 in hepatocytes. a NF-κB p65 levels in cytosolic (Cyto.) and nuclear (Nuc.) extracts in liver tissues from 10-month-old mice. b NF-κB p65 (green) immunostaining in primary hepatocytes treated with Ad-KAL for 48 h. Scale bar: 100 μm. c–f Representative immunoblot and quantification of NF-κB p65 in nuclear extracts (c), mRNA levels of TNFα (d), representative immunoblot and quantification of TNFα (e), and supernatant TNFα levels (f) in hepatocytes treated with si-Abhd5 for 48 h. g Representative immunoblot and quantification of NF-κB p65 in nuclear extracts of primary hepatocytes transfected with KAL and CGI-58 plasmid for 48 h. h NF-κB p65 (green) and CGI-58 (red) immunostaining in primary hepatocytes. Images were acquired under a laser-scanning confocal microscope. Scale bar: 25 μm. i NF-κB p65 and CGI-58 levels in Cyto and Nuc extracts of primary hepatocytes. j NF-κB p65 immunoblotting after immunoprecipitation (IP) for CGI-58 in primary hepatocytes, IP for IgG as the negative control. k The co-IP blot and quantification of CGI-58 and NF-κB p65 in primary hepatocytes treated with Ad-KAL for 48 h, IP for IgG as the negative control. l, m Oil red O staining, H&E staining and nuclear NF-κB p65 levels in liver tissues of hepatic CGI-58-overexpressing KAL-Tg mice (KAL + CGI-58LSL/+:Cre), the black arrow represents the destination blots. Scale bar for Oil red O staining: 200 μm, for H&E staining: 100 μm. Data represent the mean ± SD of three independent experiments. *p < 0.05, **p < 0.01
