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. 2021 Jan 2;473(4):647–658. doi: 10.1007/s00424-020-02510-1

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© The Author(s) 2021

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 1 — A soluble form of αKlotho downregulates SOCE without affecting Orai1/STIM1. a Representative trace showing the effect of the Klotho family on SOCE. Flag-Orai1 and YFP-STIM1 co-transfected with Klotho isoforms: αKlotho (KLA), βKlotho (KLB), or γKlotho (KLG) in HEK293FT cells. Empty vector (pEF1 vector) used as transfection control. b Quantification of peak SOCE values is expressed as mean ± SEM (n = 55–173 each group). c Immunoblotting showing transfection of Klotho isoforms and Flag-tagged Orai1 and YFP-tagged STIM1. d Quantitative real-time PCR for relative mRNA expression of Orais and STIMs in HEK293FT cells. e Representative trace of endogenous SOCE in HEK293FT cells transiently expressing full-length (KLFL) and secreted (KLΔTM) form of αKlotho. Empty vector (pEF1 vector) was used as a transfection control (Vector). f Summary of the SOCE in panel e (n = 123–186 each group). g Effect of αKlotho (KLFL and KLΔTM) on endogenous Orai1 and STIM1 protein expression in HEK293FT cells. **Denotes p < 0.01. Data were analyzed by one-way ANOVA (b left panel in d and f) and t test (right panel in d)