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. 2017 Dec 1;7:16782. doi: 10.1038/s41598-017-16977-z

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© The Author(s) 2017

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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Diabetes induced persistent activation of TGF-β₁ and Shh signalling. (a–c) Immunohistochemistry for kidney levels of (a) TGF-β₁, (b) Shh and (c) Smo at 3 and 5 weeks after IRI. Scale bar = 80 µm. (d) The kidney levels of expressed TGF-β₁ and Smad2 were measured by western blot at 5 weeks after IRI. (e) The kidney levels of expressed Shh and Smo were measured by western blot at 5 weeks after IRI. (f) The kidney levels of expressed α-SMA and fibronectin were measured by western blot at 5 weeks after IRI. Each protein expression was normalized by GAPDH. The fold-change of each protein was calculated as the ratio of averages versus non-diabetic sham. (g–l) mRNA expression of the major TGF-β₁ and Shh signalling pathway factors in the kidney were determined via qRT-PCR at 3 and 5 weeks after IRI. The targets of mRNA were (g) TGF-β₁, (h) CTGF, (i) collagen I-a1, (j) Shh, (k) Gli-1 and (l) Snail1. Each value of the target mRNA was normalized by 18 S. Values are expressed as the mean ± S.E.M. ^* p < 0.05 versus sham, ^# p < 0.05 versus non-diabetic ^† p < 0.05 versus at 3 weeks.