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. 2019 Oct 17;2:380. doi: 10.1038/s42003-019-0615-z

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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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Fig. 5 — High store-operated calcium entry in the absence of spatacsin increases cytoplasmic calcium levels. a Quantification of cytosolic calcium levels in Spg11^+/+ and Spg11^−/− fibroblasts in normal medium or medium supplemented with EGTA to lower the extracellular calcium to 0.4 mM. The graphs represent the mean ± SEM. N > 159 cells from three independent experiments. Two-way ANOVA: ***p < 0.0001. b Downregulation of STIM1 strongly abrogates store-operated calcium entry in Spg11^+/+ and Spg11^−/− fibroblasts. The graphs show the mean ± SEM. N > 55 cells from three independent experiments. Insert: western blot showing the downregulation of STIM1 in Spg11^+/+ and Spg11^−/− fibroblasts transfected with siRNA directed against STIM1. c Downregulation of STIM1 decreases the levels of cytosolic calcium in Spg11^−/− fibroblasts to those measured in Spg11^+/+ fibroblasts. The graph shows the mean ± SEM. N > 190 cells analyzed in three independent experiments. Two-way ANOVA: *p < 0.05. d Treatment of Spg11^+/+ or Spg11^−/− fibroblasts with methyl-β-cyclodextrin (MBCD) loaded with cholesterol for 1 h restores normal cytosolic calcium levels in Spg11^−/− cells. The graph shows the mean ± SEM. N > 70 cells from three independent experiments. Two-way ANOVA: **p = 0.0017, ***p = 0.0006