Volume 93, Issue 1, (January 2018)

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Presentation transcript:

Volume 93, Issue 1, (January 2018) Zhang L, Zhang Q, Liu S, et al. DNA methyltransferase 1 may be a therapy target for attenuating diabetic nephropathy and podocyte injury. Kidney Int. 2017;92:140–153 Kidney International Volume 93, Issue 1, (January 2018) DOI: 10.1016/j.kint.2017.11.001 Copyright © 2017 International Society of Nephrology Terms and Conditions

Figure 5 Expression of DNA methyltransferases (Dnmts) in vivo and in vitro and effects of DNA methylation inhibition and Sp1 and p65 knockdown on the expression of Dnmt1 in podocytes under diabetic conditions. (a) The relative mRNA levels of Dnmt1, Dnmt3a, and Dnmt3b in the renal cortex of nondiabetic BKS mice and db/db mice with or without 5-azacytidine. (b) The protein level of Dnmt1 was analyzed in the renal cortex using Western blot analysis (n = 3). (c) Densitometry analysis of 3 repetitions of Dnmt1 protein in the kidney. (d) The protein expression of Dnmt1 was analyzed in glomerular podocytes of db/db mice using immunofluorescent analysis. Double immunofluorescent staining of Dnmt1 (red), Wilms tumor 1 (WT1)–identified podocytes (green), and merged images (yellow) in the kidney from nondiabetic or diabetic mice. Bars = 20 μm. (e) The protein expression of Dnmt1 was analyzed in glomerular podocytes of 4 patients with diabetic nephropathy (DN). Representative double immunofluorescent staining of Dnmt1 (red), synaptopodin-identified podocytes (green), and merged images (yellow) in the kidney from patients with DN or control samples (normal renal tissues from patients with renal cell carcinoma). Bars = 20 μm. Quantitative immunofluorescence analysis of 20 glomeruli from 4 patients with DN demonstrated that the expression of Dnmt1 protein markedly increased in glomerular podocytes. (f) The relative expression of Dnmt1, Dnmt3a, and Dnmt3b mRNA in cultured podocytes treated with high glucose (HG) for 24, 48, and 72 hours. (g) The relative expression of Dnmt1 mRNA and the effect of DNA methylation inhibition and Sp1 or p65 knockdown on the expression of Dnmt1 in podocytes treated with HG for 48 hours. (h) Double immunofluorescent staining of Dnmt1 (red), synaptopodin (green), 4,6-diamino-2-phenylindole (DAPI)–stained nuclei (blue), and merged images in cultured podocytes treated with HG for 48 hours. Bars = 50 μm. (i) The protein level of Dnmt1 was analyzed in podocytes under different conditions using immunoblotting. (j) Densitometry analysis of 3 repetitions of Dnmt1 protein in podocytes under different conditions. All values are expressed as means ± SEM. *P < 0.05 versus BKS + PBS or normal glucose (NG); #P < 0.05 versus db/db + PBS or HG. To optimize viewing of this image, please see the online version of this article at www.kidney-international.org. Kidney International 2018 93, DOI: (10.1016/j.kint.2017.11.001) Copyright © 2017 International Society of Nephrology Terms and Conditions