Fig. 6

Nurr1 promotes the transcription of pre-miR-30e-5p, and the targeting relationship between miR-30e-5p and NLRP3. A Schematic representation illustrating the binding region of Nurr1 specific to the promoter of pre-miR-30e-5p. Nurr1 transcription factor binding sites were identified within 404–448 bp upstream of pre-miR-30e-5p in mice. B ChIP-PCR assay conducted in BV2 cells using specific primers for the promoter regions (Pro1, − 448 to − 542 bp; Pro2, − 402 to − 448 bp; Pro3, − 388 to − 402 bp; Pro4, − 216 to − 388 bp) of miR-30e-5p. C Recruitment of Nurr1 to the Pro2 region was significantly enhanced compared to the IgG group. ****p < 0.0001, Unpaired Student T test. D A dual luciferase reporter assay was employed to detect Nurr1’s transcription activity with miR-30e-5p. ****p < 0.0001, Two-way ANOVA Tukey test. E Schematic diagram of binding sites between miR-30e-5p, human and mouse NLRP3 3ʹUTR predicted by bioinformatics databases. F The targeting relationship between miR-30e-5p and NLRP3 was verified using a dual-luciferase reporter gene assay in 293T cells. 3ʹUTR-NLRP3-WT refers to NLRP3 wild type 3’UTR and 3ʹUTR- NLRP3-Mut refers to NLRP3 mutant 3ʹUTR. ****p < 0.0001, Two-way ANOVA Tukey test. G The BV2 cells were treated with miR-30e-5p mimic, miR-30e-5p inhibitor and miR-NC for 24 h. Representative immunoblots and quantitative analysis of NLRP3 protein level (H), and the relative expression level of NLRP3 mRNA (G) are shown. Gapdh was used as a loading control. G The transfection efficiency of miR-30e-5p mimic and miR-30e-5p inhibitor in BV2 cells. *, p < 0.05, ***, p < 0.001; ****, p < 0.0001, relative to the blank BV-2 cells (Ctrl); ^#, p < 0.05, ^###, p < 0.001; ^####, p < 0.0001, relative to the miR-NC. One-way ANOVA Tukey test. These experiments were repeated three times independently