Fig 1: Mesaconate suppresses cardiac ischemia/reperfusion injury by reducing macrophage inflammation through GPR35. A mRNA expression levels of TNF-α, IL-1β, IL-6, and CXCL9 in cardiac tissue (n = 5). B Secreted protein levels of TNF-α, IL-1β, IL-6, and CXCL9 in plasma (n = 5). C Immunofluorescence staining and quantification of IL-6 positive macrophages, IL-6 (Red), CD68 (Green) and DAPI (Blue) (n = 5), scale bar = 50 μm. D mRNA expression levels of TNF-α, IL-1β, IL-6, and CXCL9 in cardiac tissue (n = 4). E Secreted protein levels of TNF-α, IL-1β, IL-6, and CXCL9 in plasma (n = 5). F Immunofluorescence staining and quantification of IL-6 positive macrophages, IL-6 (Red), CD68 (Green) and DAPI (Blue) (n = 5), scale bar = 50 μm. Statistical significance was evaluated via two-tailed unpaired Student’s t-test (C, F), two-way ANOVA followed by the Bonferroni multiple-comparison test (A, B, D, E). All data were presented as mean ± SD. *p < 0.05, **p < 0.01, ***p < 0.001, n.s. no significant difference
Fig 2: Mesaconate alleviates cardiac ischemia/reperfusion injury via GPR35. A Body weight changes in mice. B Representative M-mode images. C Quantitative analysis of left ventricular ejection fraction and fractional shortening (n = 5). D Representative TTC staining images of heart sections. E Quantification of infarct area (n = 4). F Plasma levels of LDH and α-HBDH (n = 5). G Plasma levels of CK-MB and cTnI (n = 5). H Representative fluorescence images of cardiac tissue sections, ACTN2 (red), TUNEL (white), and DAPI (blue), scale bar = 50 μm. I Quantification of TUNEL-positive cells (n = 5). J Representative Western blot results of Caspase3 and Caspase7. K Quantification of (J) (n = 5). Statistical significance was evaluated via two-way ANOVA followed by the Bonferroni multiple-comparison test (C, E–G, I, K). All data were presented as mean ± SD. *p < 0.05, **p < 0.01, ***p < 0.001, n.s. no significant difference
Fig 3: Mesaconate alleviates LPS-induced macrophage ferroptosis via GPR35. A SOD activity, GSH content, LPO and MDA levels in BMDMs (n = 3). B Representative fluorescence images of BODIPY 581/591 C11 staining in BMDMs, scale bar = 50 μm. C Representative images of FerroOrange staining in BMDMs, scale bar = 50 μm. D Quantification of (B) (n = 3). E Quantification of (C) (n = 3). F SOD activity, GSH content, LPO and MDA levels in BMDMs (n = 3). G Representative images of FerroOrange staining in BMDMs, scale bar = 50 μm. H Representative fluorescence images of BODIPY 581/591 C11 staining in BMDMs, scale bar = 50 μm. I Quantification of (H) (n = 3). J Quantification of (G) (n = 3). Statistical significance was evaluated via two-way ANOVA followed by the Bonferroni multiple-comparison test (A, D–F, I, J). All data were presented as mean ± SD. *p < 0.05, **p < 0.01, ***p < 0.001, n.s. no significant difference
Fig 4: Mesaconate suppresses macrophage inflammation through GPR35. A Heat map of differentially expressed genes in mesaconate-treated BMDMs (n = 4). B Bubble plot of Gene Ontology (GO) enrichment analysis. C COG function classification of Consensus Sequence. D mRNA expression levels of GPR35 in BMDMs (n = 3). E Representative Western blot results of GPR35 after mesaconate treatment. F Representative Western blot results of GPR35. G mRNA expression levels of TNF-α, IL-1β, IL-6, and CXCL9 in LPS-stimulated BMDMs treated with increasing doses of mesaconate (n = 3). H Secreted protein levels of TNF-α, IL-1β, IL-6, and CXCL9 in culture supernatants (n = 3). I Immunofluorescence staining and quantification of p-NF-κB in BMDMs (p-NF-κB (Yellow) and DAPI (Blue) (n = 3), scale bar = 50 μm. Statistical significance was evaluated via two-tailed unpaired Student’s t-test (D), two-way ANOVA followed by the Bonferroni multiple-comparison test (G–I). All data were presented as mean ± SD. *p < 0.05, **p < 0.01, ***p < 0.001, n.s. no significant difference
Supplier Page from CUSABIO Technology LLC for Customized Mouse Gpr35 Protein (in vitro E.coli)