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. 2019 Aug 6;9:11357. doi: 10.1038/s41598-019-47778-1

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© The Author(s) 2019

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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miRNA 27b-3p and miRNA 1228-3p validation on kidney tissues. (A) qPCR of miRNA 1228-3p (upper panel) and miRNA 27b-3p (lower panel) expression levels on RNA extracted from kidney biopsies of 5 patients with normal kidney morphology (NK), 12 DN and 8 T2D-MN patients. Center lines show the medians; box limits indicate the 25^th and 75^th percentiles as determined by R software; whiskers extend 1.5 times the interquartile range from the 25^th and 75^th percentiles, outliers are represented by dots. (B) In situ Hybridization of miRNA 1228-3p and 27b-3p expression levels on kidney biopsies of DN and T2D-MN patients. snU6: small nuclear RNA U6 positive control. Scramble: DIG-labeled Scramble-miR probe negative control. ISH quantification was performed as described in methods. The histogram represents the mean ± SD. (C) Immunohistochemical analysis on kidney biopsies of patients with T2D and membranous nephropathy (MN) or diabetic nephropathy (DN).