Fig 1: Effect of neutralizing antibodies for chemokines/chemoattractans in monocytes/MΦs in vitro cell migration assays. (A) The effect of the neutralizing antibodies against CCL2 (500 ng/mL), CCL3 (500 ng/mL), and CCL21 (500 ng/mL) in the monocytes/MΦs migration assay under the chemoattractant effect of the CM of the glomeruli from DN rats. HAM-F10 and 5% FBS were used as negative and positive control, respectively, of in vitro stimuli for migration. DAPI was used to stain the nuclei of monocytes/MΦs. Representative quadrants show monocytes/MΦs in blue (400× magnification). (B) Quantification of positive intraglomerular area (%) of deconvoluted color for DAB in (A). Graphs represent distribution of each sample and the mean ± S.D. * p < 0.05, and *** p < 0.001 versus Ctrl rats. # p < 0.05 and ### p < 0.001, DN + MRS1754 versus DN rats. n = 4.
Fig 2: Expression of chemokines/chemoattractans in glomeruli of DN + MRS1754 rats. (A) Protein expression of CCL3 and CCL21 (green color) by immunohistofluorescence in glomeruli (red color) of Ctrl, DN, and DN + MRS1754 rats. The dotted lines represent glomeruli areas. (B) Quantification of positive intraglomerular area (%) of (A). Graphs represent the mean ± S.D. * p < 0.05, ** p < 0.01 and *** p < 0.001 versus Ctrl rats. # p < 0.05 and ## p < 0.01, DN + MRS1754 versus DN rats. n = 4.
Fig 3: A2BAR antagonism attenuates inflammatory mediators in diabetic rats. (A) The graphs depict the levels of the chemokines MCP-1 and CCL3 released from glomeruli and the urinary TGF-β levels in the rat experimental groups. n = 6 samples per duplicate. *, p < 0.05 vs. control; #, p < 0.05 vs. DN; &, p < 0.05 vs. control. (B) The heat map highlights changes in the transcript levels of selected inflammation-related genes in the experimental rat groups. The table shows values related to a decrease in the transcripts in the DN + MRS1754 group vs. the DN group.
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