Fig. 2

P65 is required for LPS-induced inflammasome activation and cell movements. a LPS induced P65 phosphorylation and transportation to nucleus. b Cells were transfected with siNC (control) or siP65 (knockdown of P65 with specific siRNA, 50nM for 48 h), then treated with 1μg/ml LPS for 24h. Phosphorylated P65, nuclear P65, and inflammasome activation makers Caspase-1-P20 and IL-1β-P17 were determined by western blotting. c To inhibit NF-κB activity, 50μM Bay11-7082 was added 3h prior to the addition of LPS. DMSO was solvent of Bay11-7082. Phosphorylated and nuclear P65 and inflammasome activation makers were determined by western blotting. d Cells were transfected with siNC (control) or siP65 for 48 h, then treated with LPS 1μg/ml for 24 h. Enzyme activity of caspase-1 was detected by Caspase-Glo^® 1 Inflammasome Assay Kit. e Transwell assay was performed to investigate migration and invasive properties of CRC cells with or without LPS treatment in the presence or absence of siP65 transfection. Data are presented as mean±SEM. *P<0.05, **P<0.01, ***P<0.001. N, nuclear; p-P65, phosphorylated P65; RLU: relative luminometer units