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. 2018 Feb 19;8:3249. doi: 10.1038/s41598-018-21405-x

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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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SPAK inactivation in WNK1^+/FHHt mice decreases NCC abundance and phosphorylation below control levels. Upper panel: Representative immunoblots of NCC and phosphorylated NCC-Thr53 performed on the membrane-enriched fractions of the renal cortex of mice of each genotype. Lower panel: Densitometric analysis. NCC abundance and phosphorylation are increased in WNK1^+/FHHt mice compared to control mice. NCC abundance and phosphorylation in WNK1^+/FHHt:SPAK^243A/243A and SPAK^242A/243A mice are corrected below the control levels. Number of animals: 4 control, 5 WNK1^+/FHHt, 4 WNK1^+/FHHt:SPAK^243A:243A and 4 SPAK^243A:243A male mice. Values are means ± s.e.m. ^ap < 0.05 vs. controls. ^bp < 0.05 vs. WNK1^+/FHHt. ^cp < 0.05 vs. SPAK^243A/243A-WNK1^{+/FHHt FHHt} (Kruskal-Wallis one-way analysis of variance followed by Mann-Whitney test). Immunoblot images are cropped images. The full-length images can be found in Supplementary Figure S3.