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. 2017 Oct 11;7:12981. doi: 10.1038/s41598-017-12819-0

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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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Mutation of K706, K828, and K909 residues in NCC increases membrane abundance of pT58-NCC when expressed in MDCKI cells. (A) Representative immunoblots of pT58-NCC and NCC in immunoprecipitated (anti-FLAG-tag) samples isolated from the apical biotinylated pool of various MDCKI cells under control (Ctrl) or low chloride (LC) conditions. (B) Semi-quantitative assessment of pT58-NCC levels at the plasma membrane under Ctrl conditions. Data were analyzed using one-way ANOVA followed by Tukey-Kramer multiple comparison test and presented as means ± S.E.M. (n = 6–9) *indicates significant difference compared to WT-NCC under Ctrl conditions. (C) Semi-quantitative assessment of pT58-NCC levels at the plasma membrane under Ctrl or LC conditions. Data were analyzed using a two-way ANOVA followed by Tukey-Kramer multiple comparison test. Data are means ± S.E.M. (n = 6–9) *indicates significant difference between LC and Ctrl conditions for individual cell line. ^#Represents significant difference compared to WT-NCC following LC stimulation.