Fig. 3

NF-κB binds to the promoter region of NEAT1 after LPS-stimulated p65 nucleus translocation (a) A schematic diagram of potential p65 binding element (two possible binding sites) in the promoter region of NEAT1 predicted by Jaspar database. A wt-NEAT1 promoter luciferase reporter vector and a mut-NEAT1 promoter luciferase reporter vector were constructed by mutating any or both of the predicted binding sites. b The indicated vectors were transfected into HEK293 cells and then treated with PBS or LPS (20 ng/ml for 4 h); the luciferase activity was determined. c-d The real-time ChIP assay showed that the level of p65 antibody binding to NEAT1 promoter was much greater than that of IgG. e A549 and H1299 cells were transfected with si-p65 or p65 vector; the protein levels of p65 and p-p65 were determined using Immunoblotting in the presence or absence of LPS (20 ng/ml for 4 h). f The mRNA expression of NEAT1 was determined using real-time PCR assays. The data are presented as mean ± SD of three independent experiments. *P < 0.05, **P < 0.01, compared to PBS group; #P < 0.05, ##P < 0.01, compared to si-NC or vector group