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. 2019 Feb 4;9:1175. doi: 10.1038/s41598-018-37602-7

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© The Author(s) 2019

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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SPCA2 regulates Kv10.1 and Orai1 total and membrane expression. (A) Representative western blots (a) of Kv10.1 and Orai1 proteins levels in siCtl and siSPCA2 MCF-7 cells after 48 h of starvation treated or not with collagen 1. Results were normalized as a percentage of the untreated control condition (−Coll). Values are reported as means of all the experiments. (b) Histograms representing the averages ± standard error of results obtained by western blots. N = 5, **p < 0.01; ANOVA followed by Holm-Sidak post hoc tests. (B) Effect of SPCA2 silencing on Kv10.1 and Orai1 membrane expression. Representative western blots showing membrane fraction enrichment of Kv10.1 (a) and Orai1 (b) proteins in siCtl and siSPCA2 MCF-7 cells after 48 h of starvation. The membrane fractions were obtained through a biotinylation protocol. Results were normalized as a percentage of the siCtl collagen 1 treated condition. Values are reported as mean of all the experiments. N = 3. (C) Whole cell currents (a) recorded in siCtl and siSPCA2 MCF-7 cells after 48 h of starvation treated with collagen 1. 500 msec voltage ramps from −100 to +100 mV from a holding potential of −40 mV were applied to record Kv10.1 channel activity. Values are reported as mean ± standard error. (b) Histograms representing the averages ± standard error of currents values at +100 mV. siCtl (n = 10), siOrai1 (n = 5); *p < 0.05; Two-samples t-test.