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. 2018 Apr 26;8:6630. doi: 10.1038/s41598-018-24340-z

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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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Tissue fibrosis was assessed by Masson’s Trichrome staining, Col-I and Col-IV levels using immunohistochemistry. Masson’s trichrome staining was performed on kidney sections at 28 days post contrast (C) or saline (S) administration. A representative image of Masson’s Trichrome staining on day 28 is shown (A). There is increased tubular fibrosis (blue color) in animals with contrast (C) administration compared to saline (S). Representative images of collagen-1 (B) and collagen-IV (D) staining at 7 (D7) and 28 (D28) are depicted. The percentage of brown staining over tissue area was calculated using Zen 2.3 software (C and E). There was significantly higher collagen-I (C) and collagen-IV (E) staining in kidney with C administration compared to S. All images are at 40X magnification. Each bar in the bar diagram represents mean ± SEM of 4–6 animals per group. Two-way ANOVA with Student t test was performed with Bonferroni correction was performed. Significant differences between the respective time points among C and S indicated by * at P < 0.05.