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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 NCC at specific lysines influences lysosomal or proteasomal NCC degradation when expressed in MDCKI cells. (A) Representative immunoblots of NCC in lysates isolated from various MDCKI cell lines treated with vehicle (ethanol), 50 µM cycloheximide, 50 µM cycloheximide plus 20 µM MG132, 50 µM cycloheximide plus 400 µM chloroquine, 50 µM cycloheximide plus 20 µM MG132 and 400 µM chloroquine for indicated time. (B) Semi-quantitative analysis of NCC levels in various MDCKI cell lines following the different treatments. NCC levels in cells expressing K706R and K909R mutants are significantly different that WT-NCC expressing cells. Data were analyzed using two-way ANOVA followed by Tukey-Kramer multiple comparison test and presented as means ± S.E.M. (n = 6–9). *Represents significant change compared to WT-NCC under similar conditions.