Fig 2: Nur77 deficiency exacerbates NLRP3 inflammasome-mediated inflammation in PMs. PMs from WT or Nur77-/- mice were stimulated with or without ox-LDL (100 µg/ml) for 24 h and then were collected for analysis. (a) The LDH activity in supernatant was assessed by LDH Cytotoxicity Assay Kit. The expression levels of NLRP3 inflammasome-related proteins NLRP3 (b), c-caspase-1 (c), c-IL-1ß (d), and GSMD-N (e) were determined by Western blotting. Relative protein levels of NLRP3 (f), c-caspase-1 (g), c-IL-1ß (h), and GSMD-N (i). (j) The mice NLRP3 promoter reporters, blank PGL3, and pcDNA or Nur77 plasmid were transiently transfected into Raw264.7 cells; after 24 h, the dual-luciferase activity was measured. (k) Raw264.7 cells were treated with or without ox-LDL (100 µg/ml) for 24 h; DNA fragments from the Raw264.7 cells that contain regions of the NLRP3 promoter were immunoprecipitated with the anti-Nur77 antibody. The expression was identified by q-PCR. n = 3. Values are shown as the mean ± SEM. *P < 0.05.

Fig 3: NR4A1 is associated with LEF1 expression in PTC.A GSEA analysis of GSE27155 by Oncogenic Pathway database to identify the potentially enriched signatures in higher NR4A1 expression of PTC tissues. B The heatmap showed the RT-qPCR results of LEF1 target gene expression in NR4A1 knockout and control BCPAP and TPC-1 cells. Unpaired Student’s t test, *P < 0.05, **P < 0.01, ***P < 0.001. C Western blot analysis of LEF1 protein levels in NR4A1 knockout and control cells. D RT-qPCR results of LEF1 mRNA levels in NR4A1 knockout and control cells. The NR4A1 knockout group was compared to the control group, Student’s t test, ***P < 0.001. E Representative IHC stainings of NR4A1 (upper) and LEF1 (lower) proteins in PTC tumorous tissues with high or low NR4A1 expression. Scale Bars = 50 µm. F Statistical analysis was conducted based on the NR4A1, and LEF1 protein expression in the TMA with 42 cases of PTC. Chi-square test. G Western blot analysis to validate LEF1 knockout efficiency in BCPAP and TPC-1 cells. H Cell viability assay via CCK8 method to compare the proliferation rate between LEF1 depleted and control BCPAP and TPC-1 cells. One-way ANOVA test, **P < 0.01, ***P < 0.001. I Representative images and semiquantitative analyses of IHC staining of LEF1 protein in a TMA of paired PTC tumorous tissues and adjacent normal tissues. Chi-square test, **P < 0.01. Scale bars = 500 µm. J The heatmap illustrated the association of different clinical characters in PTC patients with high or low LEF1 expression. Statistical significance was assessed by the Chi-square test.

Fig 4: Effect of Nur77 deficiency on macrophage inflammatory responses of atherosclerotic lesions. (a) Representative immunofluorescence staining of CD68 (red), c-caspase-1 (green), and their colocalization (yellow) of carotid atherosclerotic lesions in mice from each group (scale bar = 50 µm). (b) Quantification of c-caspase-1+ CD68+ areas relative to the total CD68+ area (n = 8). (c) Representative immunofluorescence staining of CD68 (red), c-IL-1ß (green), and their colocalization (yellow) of carotid atherosclerotic lesions in mice from each group (scale bar = 50 µm). (d) Quantification of c-IL-1ß+ CD68+ areas relative to the total CD68+ area (n = 8). Values are shown as the mean ± SEM. *P < 0.05.

Fig 5: Csn-B activated Nr4a1 and attenuated cocaine behavior.a Timeline of Csn-B administration (10 mg/kg) acute and repeated (2× daily, 4 days). b Nr4a1 was activated after acute and repeated Csn-B treatment (n = 5 mice/group; two-way ANOVA, followed by Bonferroni’s multiple comparisons test, 6 h P = 0.028, 1-day P = 0.0174, 4-days P = 0.0168), normalized to GAPDH, relative to DMSO controls. *P < 0.05. c Csn-B administration activated Cartpt after acute but not following repeated Csn-B administration (two-way ANOVA, followed by Tukey’s multiple comparisons test, 6 h P = 0.0307, 1-day P > 0.9999; 4-day P = 0.0978), normalized to GAPDH, relative to DMSO controls. *P < 0.05. d Acute Csn-B administration enriched H3K27me3 at the Cartpt promoter (n = 6 mice/group; two-way ANOVA, followed by Tukey’s multiple comparisons test, 6 h P = 0.0276), normalized to input, relative to DMSO controls. *P < 0.05. e There were no significant change in H3K27ac enrichment following acute and repeated Csn-B administration (n = 6 mice/group; two-way ANOVA, followed by Tukey’s multiple comparisons test, 6 h P = 0.0276), normalized to input, relative to DMSO controls. *P < 0.05. f Acute Csn-B administration enriched H3K4me3 at the Cartpt promoter (n = 6 mice/group; two-way ANOVA, followed by Tukey’s multiple comparisons test, 6 h P = 0.033), normalized to input, relative to DMSO controls. *P < 0.05. g Schematic of cocaine conditioned place preference (CPP) paradigm and Csn-B administration time course. h Repeated Csn-B administration (green circles) attenuated cocaine CPP (n = 7 mice/group, two-way repeated measures ANOVA), relative to DMSO control. *P < 0.05. i Repeated Csn-B administration activated Nr4a1 and Cartpt following cocaine CPP (n = 7–8, unpaired two-tailed t-test, Nr4a1 P = 0.0157, Cartpt P = 0.005), normalized to GAPDH, relative to DMSO. *P < 0.05. j Schematic of cocaine conditioned place preference (CPP) paradigm and Csn-B administration time course. k Acute Csn-B administration inhibited cocaine CPP (n = 7 mice/group, two-way repeated measures ANOVA) relative to DMSO control. *P < 0.05. l Acute Csn-B administration activated Nr4a1 and Cartpt following cocaine CPP (n = 7, unpaired two-tailed t-test, Nr4a1 P = 0.0404, Cartpt P = 0.0075), normalized to GAPDH, relative to DMSO. *P < 0.05. All error bars represent mean +/- s.e.m. Source data and statistics provided as a Source Data file.