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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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OSR1 remains at the apical membrane of SPAK^243A/243A and WNK1^+/FHHT:SPAK^243A/243A DCT, where its phosphorylation tends to increase. Upper panel: Representative Immunoblots performed on total cortex protein extract with antibodies against total SPAK, total OSR1, pSPAK/pOSR1 (S-motif) and Cab39. Lower panel: Densitometric analysis. SPAK inactivation mutation does not affect SPAK expression and S-motif phosphorylation, while it tends to slightly increase OSR1 expression and phosphorylation. 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 mice^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 S5.