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. 2022 Aug 27;13:5062. doi: 10.1038/s41467-022-32477-9

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© The Author(s) 2022

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 3 — a–c Exemplary histological images of human kidney sections stained for p21 (top) or γ-H2A.X (histone H2A family X, bottom) obtained from non-diabetic controls (C; n = 6) or diabetic patients without (DM-DKD; n = 6) or with (DM + DKD; n = 5) DKD; p21 is detected by HRP-DAB reaction, brown; hematoxylin nuclear counter stain, blue; γ-H2A.X is immunofluorescently detected, red; DAPI nuclear counterstain, blue. Dot plots summarizing results for p21 (b) and γ-H2A.X (c); Scale bars represent 20 μm. one-way ANOVA with Sidak’s multiple comparison test. d, e Urinary p21, detected by immunoblotting (d) or ELISA (e; pg/ml) is markedly increased in patients with DKD (n = 26) compared to non-diabetic controls with normal kidney function (C; n = 22). Urinary p21 levels in patients with other causes of chronic kidney disease (oCKD; n = 18) are comparable to those in patients with DKD. Exemplary immunoblot (d) and dot plots reflecting mean ± SD showing the distribution of the urinary levels of p21 (e, Kruskal–Wallis test with Dunn’s multiple comparison test. f Violin plots showing the distribution of the urinary levels of p21 (pg/ml; ELISA) in normoglycemic controls (C; n = 36) and diabetic individuals which are classified according to KDIGO criteria from the LIFE-adult cohort (low risk, n = 52; moderate risk, n = 53; high risk, n = 29 and very high risk, n = 18). Kruskal-Wallis test with Dunn’s multiple comparison test. g Receiver operating characteristic (ROC) analyses of urinary p21 (pg/ml; ELISA) in diabetic individuals with low or moderate risk of CKD (blue) and in diabetic patients with high or very high risk of CKD (red) compared to non-diabetic controls (C). AUC: area under the curve. h–j Negative correlation of urinary p21 (pg/ml, ELISA) with estimated glomerular filtration rate (eGFR, ml/min/1.73 m², (h) and positive correlation with urinary albumin creatinine ration (UACR, mg albumin /g creatinine, (i) or with cystatin C serum levels (Cyt. C, mmol/l, j) in diabetic individuals from the LIFE-adult cohort. Pearson’s correlation. k–n line-graphs illustrating individual changes of urinary p21 (k; ELISA, pg/ml, m; immunoblot) or HbA1c (l, n) in diabetic patients with known DKD (n = 10) before (−) and after (+) SGLT2i treatment (k, l) or before (−) and after (+) fasting mimicking diet (FMD, m, n). Paired t-test. Gels/blots were processed in parallel; source data are provided as a Source Data file-Fig-3.