Fig. 5

MALAT1 positively regulated NLRP3 expression level through binding to STAT1. A The protein bands of sense and anti-sense MALAT1 groups after RNA-pull down assay was performed. STAT1 was marked in the image. B RNA pull-down assay for STAT1 in input, anti-sense and sense MALAT1 groups. C MALAT1 enrichment of input, IgG and anti-STAT1 groups were detected by RIP assay. The MALAT1 relative enrichment was normalized on input. D FISH result exhibited the subcellular localization of MALAT1 (red) and STAT1 (green) of the BV2 cells. The cells were hybridized with the Cy-2 labeled anti-STAT1 and Cy-7 labeled MALAT1 probes. E The potential binding site sequences of STAT1 with NLRP3 (upper). DNA binding motif of STAT1 predicted by Jaspar (lower). F Relative luciferase activities of the BV2 cells were detected 48 h after transfection of luciferase reporter vectors. G Relative luciferase activities of the BV2 cells were detected 48 h after they were transfected with wild and mutant types STAT1 luciferase reporter vectors. H The affinity of STAT1 in the promoter of NLRP3 detected using ChIP assay. I NLRP3 levels in the the BV2 cells were measured by RT-qPCR after MALAT1 knockdown or overexpression. J Western blot for NLRP3 of the BV2 cells transfected with indicated ad-shRNA or overexpression plasmids. (n=3), ^**P<0.01, NC, negative control. Cite 1, the sequence of NLRP3 promoter regions 277–287; cite2, the sequence of NLRP3 promoter regions 1072–1082; cite2, the sequence of NLRP3 promoter regions 1238–1248