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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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Contrast exposure increases pSMAD3 in vitro and in vivo. Western blot analysis of pSMAD3 in HK-2 cells treated with contrast at indicated time points (A) and image quantification (B). There is a significant increase in the mean pSMAD3/SMAD3 levels in the HK-2 cells treated with contrast compared to baseline (Ctrl) at 1, 3, and 12 hours (P < 0.05). (C) is a representative image of immunostaining for pSMAD3 in kidney sections with contrast (C) administration or saline (S) at day 2 (D2) or day 7 (D7). All images are at 40X magnification. Arrowhead indicates brown nuclei positive, which are for pSMAD3. The percentage of pSMAD3 positive nuclei over total number of nuclei was presented in the bar graph (D). Each bar represents mean ± SEM of 4–6 animals per group. Two-way ANOVA with Student t test with Bonferroni correction was performed. Significant differences between the respective time points among C and S indicated by * at P < 0.05.