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. 2018 Dec 21;8:18038. doi: 10.1038/s41598-018-36061-4

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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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emx1 deficiency is not associated with changes in cell death or proliferation within the pronephros. (A) Confocal imaging of double FISH analysis (lateral view) of 28 ss WT and emx1 deficient embryos for the renal epithelial cell marker cdh17 (green), the pan-distal segment marker clcnk (red), combined with protein localization of active caspase-3 (grey) and DAPI staining (blue) through IHC. Yellow boxes demarcate areas shown in insets. Scale bar = 50 μm. (B,C) Quantification of caspase-3⁺ cell number and clcnk domain. (D) Confocal imaging of FISH analysis (lateral view) of 28 ss WT and emx1 deficient embryos for the DE marker slc12a1 (green), with protein localization of pH3 (red) and DAPI (blue) using IF. Yellow lines demarcate the DE domain. Scale bar = 20 μm. (E,F) Quantification of pH3⁺ cells in the DE and average DE length. **p < 0.01; N.S. = not significant. Error bars indicate standard error.