Fig. 4

TNF-α up-regulates SUMO2/3 and promotes p65 SUMOylation. a-b SMMC7721 cells were treated with TNF-α (10 ng/ml) for 30 min (a) and 8 h (b), respectively. The nuclear (N) and cytoplasmic (C) fragments were extracted by using the protein extraction kit according to the manufacturer’s instruction. The proteins were blotted with the antibodies as indicated. Tubulin and histone H3 were used as the markers of cytoplasm and nucleus, respectively. c-d SMMC7721 cells were co-transfected with myc-Ubc9 and GFP-SUMO2 (c) or GFP-SUMO3 (d). At 24 h after transfection, the cells were treated with TNF-α (10 ng/ml) for 30 min. Then the cells were lyzed and incubated with 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