Fig. 3

SUMO2/3 interacts with p65 in liver tissues and hepatoma cell lines. a The similar profiles of p65 and SUMO2/3 in the liver tissues. p65 and SUMO2/3 were detected using immunohistochemistry in the serial sections of the liver tissues. The images of the rectangles in the upper panels are magnified in the lower panel. The scale bars were shown as indicated. The arrows indicate the tumor tissues. b-d Co-localization of SUMO2/3 and p65. Double-labeled immunofluorescent staining was performed using the antibodies against p65 (green) and SUMO2/3 (red) in the liver tissues (b), HepG2 cells (c) and SMMC7721 cells (d). The nuclei were stained with DAPI (blue). In panel b, the images of the rectangles in the upper panels are magnified in the lower panels. The scale bars were shown as indicated. e-f Interaction of p65 and SUMO2/3 in the hepatoma cells. SMMC7721 cells were co-transfected with myc-Ubc9 and GFP-SUMO2 (e) or GFP-SUMO3 (f). At 24 h after transfection, the cells were lyzed and incubated with Protein A agarose containing anti-p65 antibody. g Interaction of p65 and SUMO2/3 in the liver tissues. The non-tumor tissues collected from HCC patients were lyzed and co-immunoprecipitation assay was performed using anti-p65 antibody. The isotype IgG was used as a negative control. The bound proteins were blotted by using the antibodies as indicated. 2 % total lysate was loaded as input