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. 2017 Aug 29;7:9815. doi: 10.1038/s41598-017-10168-6

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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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Effects of FXR activation on hypoxia-induced oxidative stress in primary cultured proximal tubular cells (PTCs). H/R, hypoxia/re-normoxia. (A–D) Representative images of (A) TUNEL staining, (B) CellROX staining, (C) TMRM staining and (D) immunostaining for Grp78 on PTCs treated with vehicle under (a) normoxia and at 24 h after exposure to hypoxia (b) without 6-ECDCA treatment or (c) with 6-ECDCA treatment (scale bar 50 μm). Staining colors correspond to the labels on the left side of each panel. Cells were counterstained with hematoxylin (blue) or DAPI (blue). (E–H) Quantitative analysis of positive (E) TUNEL staining and (F) ROS staining per high power field and (G) ATP production and (H) intracellular MDA in PTCs. n = 4/group. Data are means ± SEM, one-way ANOVA with Bonferroni’s test. *p < 0.05. (I) Representative images of GSH detection in PTCs with different treatments (scale bar 50 μm). H/R, hypoxia/re-normoxia. Staining colors correspond to the labels on the left side of each panel. Cells were counterstained with ActinGreen (green). (J) Representative images of CellROX staining of PTCs treated as indicated (scale bar 100 μm).