Fig 1: DJ-1 regulated the disassociation of NLRX1 from TRAF6 after cerebral I/R injury via SHP-1. a, g After virus and TPI-1 were used to overexpress DJ-1 and inhibit SHP-1, respectively, Western blotting was used to detect NLRX1, TRAF6, and SHP-1 in rats. b, h After virus and TPI-1 were used to overexpress DJ-1 and inhibit SHP-1, respectively, Western blotting was used to detect NLRX1, TRAF6, and SHP-1 in astrocytes. c, i After treatment with an SHP-1 inhibitor, Western blotting was used to detect the cytokines IL-1β, IL-6, and TNF-α in rats. d, j After treatment with an SHP-1 inhibitor, Western blotting was used to detect the cytokines IL-1β, IL-6, and TNF-α in astrocytes. e Immunoprecipitation and immunoblot analyses of NLRX1-TRAF6 in rats. f Immunoprecipitation and immunoblot analyses of SHP-1-TRAF6 in rats. The data are expressed as the mean ± SEM. *p < 0.05 vs. the sham group; #p < 0.05 vs. the MCAO group; &p < 0.05 vs. the overexpression group; ★p < 0.05 vs. the DMSO group; △p < 0.05 vs. the DMSO group. n = 6 per group. The data are expressed as the mean ± SEM. *p < 0.05 vs. the control group; #p < 0.05 vs. the OGD/R group; &p < 0.05 vs. the overexpression group; ★p < 0.05 vs. the DMSO group; △p < 0.05 vs. the DMSO group. n = 6 per group
Fig 2: DJ-1 regulated SHP-1-TRAF6 and NLRX1-TRAF6 interactions after cerebral I/R injury. a, c Immunoprecipitation and immunoblot analyses of NLRX1-TRAF6 in rats. NLRX1 disassociated from TRAF6 after MCAO/R, and NLRX1 interacted with TRAF6 after treatment with DJ-1 siRNA. b, d Immunoprecipitation and immunoblot analyses of SHP-1-TRAF6 in rats. SHP-1 was associated with TRAF6 in the MCAO group, and SHP-1 disassociated from TRAF6 after treatment with DJ-1 siRNA. n = 6 per group
Fig 3: DJ-1 regulated the expression of NLRX1, TRAF6, and SHP-1 after cerebral I/R injury. a, c Western blot detecting NLRX1, TRAF6, and SHP-1 in rats. b, d Western blot detecting NLRX1, TRAF6, and SHP-1 in astrocytes. e Immunohistochemistry was used to measure NLRX1 expression in mitochondria in rats after DJ-1 knockdown. Original magnification, × 400. f Immunocytochemistry was used to measure NLRX1 expression in mitochondria in astrocytes. Original magnification, × 600. Fluorescence microscopy was used to assess NLRX1 expression, which is indicated by red fluorescence. HSP60 expression is indicated by green fluorescence. Cell nuclei were stained with DAPI. The data are expressed as the mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. n = 6 per group
Fig 4: Mechanisms by which DJ-1 regulates inflammation. DJ-1 exerts an anti-inflammatory effect by decreasing the expression of cytokines IL-1β, IL-6, and TNF-α in cerebral I/R injury. DJ-1 facilitates the dissociation of NLRX1 from TRAF6. DJ-1 upregulates SHP-1 expression and facilitates the interaction between NLRX1 and TRAF6. DJ-1 induces the dissociation of NLRX1 from TRAF6 via SHP-1
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