Journal: Nature Communications

Article Title: Loss of endothelial glucocorticoid receptor accelerates diabetic nephropathy

doi: 10.1038/s41467-021-22617-y

Figure Lengend Snippet: a Western blot analysis of vimentin, HIF1α, Snail1, active β-catenin, CPT1a, and PPARα in isolated endothelial cells from the kidneys of nondiabetic and diabetic control and GR ECKO mice. Endothelial cells were isolated and cultured from six biologically independent mice/group. Densitometry calculations from six samples/group combined from three independent experiments are shown and are normalized to β-actin or HSP90 as indicated. Representative blots are shown. Data are mean ± SEM. b Western blot analysis of αSMA, fibronectin, active β-catenin and β-catenin in isolated endothelial cells from the kidneys of diabetic control, GR ECKO , Apoe −/− , and DKO mice. Endothelial cells were isolated and cultured from six biologically independent mice/group. Densitometry calculations from six samples/group combined from three independent experiments are shown and are normalized to β-actin or HSP90 as indicated. Data are mean ± SEM. Representative blots are shown. c Immunofluorescence analysis of αSMA/CD31 and TGFβR1/CD31 was performed in the kidneys of nondiabetic and diabetic control, GR ECKO , Apoe −/− , and DKO mice. FITC-labeled αSMA, FITC-labeled TGFβR1, rhodamine-labeled CD31 and DAPI (nuclei, blue) were used. Representative merged images are shown; original magnification ×300. Scale bar 50 μm. N = 7 biologically independent mice/group. Data are mean ± SEM. For all panels, one-way ANOVA with Tukey post-test was used for the analysis of statistical significance. Significance * p < 0.05, ** p < 0.01, *** p < 0.001. Source data are provided in the source data file.

Article Snippet: Anti-TGFβR1 (ab31013), PPARα (ab215270), mouse monoclonal anti-vimentin (RV202) (ab8978), rabbit polyclonal anti-αSMA (ACTA2) (ab5694), anti-HIF1α (ab516008), and goat polyclonal anti-Snail1 (ab53519) antibodies were purchased from Abcam (Cambridge, UK).

Techniques: Western Blot, Isolation, Control, Cell Culture, Immunofluorescence, Labeling