Fig. 5

SUMO2/3 stabilizes the cytoplasmic p65. a-b Dose-dependent effects of SUMO2/3 on p65 level. SMMC7721 cells were transfected with GFP-SUMO2 (a) or GFP-SUMO3 (b). The blank vector (pEGFP-C1) was transfected to balance the total DNA. At 24 h after transfection, the cells were collected and processed for immunoblotting. The data were expressed as the means ± SD of at least three independent experiments. **p < 0.01, ***p < 0.001, compared with pEGFP-1 vector. c SUMO2/3 knockdown decreased p65 level. SMMC7721 cells were transfected with SUMO2/3-siRNA (siRNA) or negative control siRNA (control). Western blotting assay was performed 48 h after transfection. The data were expressed as the means ± SD of at least three independent experiments. *p < 0.05, compared with the control. d-e SUMO2/3 increased the cytoplasmic p65. SMMC7721 cells were transfected with GFP-SUMO2 (d) or GFP-SUMO3 (e). The blank vector (pEGFP-C1) was transfected to balance the total DNA. Nuclear and cytoplasmic proteins were extracted by using the protein extraction kit according to the manufacturer’s instructions. Tubulin and histone H3 were used as the markers of cytoplasm and nucleus, respectively. f SUMO2/3 knockdown decreased the cytoplasmic p65. SMMC7721 cells were transfected with SUMO2/3-siRNA. NC (negative control) siRNA was used as a control. Nuclear and cytoplasmic proteins were extracted 48 h after transfection and processed for Western blotting