Fig 2: Effect of Nur77 deficiency on the cellular composition of atherosclerotic lesions in ApoE-/- mice. Representative images of carotid atherosclerotic lesion composition in mice from each group by staining with Oil red O, smooth muscle cell a-actin (SMC a-actin), CD68, or collagen. Quantification of Oil red O+, SMC a-actin+, CD68+, and collagen+, areas relative to the total plaque area. Scale bar = 100 µm; n = 8. Values are shown as the mean ± SEM. *P < 0.05.

Fig 3: 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 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: Nur77 deficiency promotes atherosclerotic plaque progression and destabilization. (a) Schematic illustration of the study design of the timeline and treatment protocol. (b) Representative magnetic resonance images of the cross-sections of LCCA in mice from each group 4 weeks after the surgeries (scale bar = 2 mm). The vascular lumen was shown by a white arrow. (c) Representative gross images of LCCA segments (scale bar = 2 mm) and HE staining of LCCA sections in mice from each group (scale bar = 100 µm). (d) Quantification of the carotid atherosclerotic plaque areas in mice from each group (n = 12–17). Values are shown as the mean ± SEM. LCCA: left common carotid artery; LRA: left renal artery; L: left. *P < 0.05.