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. 2018 Jul 12;8:10548. doi: 10.1038/s41598-018-28439-1

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© The Author(s) 2018

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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Effects of probucol on the changes of both Smad1 and Smad3 and their phosphorylation forms in diabetic mice. (a) Immunohistochemical staining of Smad1, pSmad1C, pSmad1L, Smad3, pSmad3C and pSmad3L in glomeruli of three groups treated with probucol (normal control, db/db and Smad3^+/−; db/db mice). Each lane represents representative data. (b) Quantitation of Smad1, pSmad1C, pSmad1L, Smad3, pSmad3C, and pSmad3L expression in the above three groups. Smad1 and Smad3 were assessed by quantitative PCR analysis, and normalized to the expression of Rn18s. As to pSmad1C, pSmad1L, pSmad3C, and pSmad3L, the number of each phosphorylated Smad1 or Smad3 protein positive nuclei was counted, and the mean values (per one glomerulus) were calculated (results are expressed as the mean ± S.E., NS, not significant, *p < 0.05 versus probucol-treated normal control mice, **p < 0.05 versus probucol-treated db/db mice, t test). C, D, and S3D stand for normal control mice, db/db mice, and Smad3^+/−; db/db mice, respectively.