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. 2019 Dec 3;9:18152. doi: 10.1038/s41598-019-54665-2

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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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Contractility analysis of EHTs under electrical stimulation. EHTs were exposed to Tyrode’s solution containing different external [Ca²⁺] (0.1–1.8 mM) and paced at a murine physiological frequency (6 Hz) at culture days 14–16. (a) Maximal force of contraction of the different groups at 1.8 mM external [Ca²⁺] and (b) corresponding contraction time (T1_20%) and (c) relaxation time (T2_20%). (d) Relative force-[Ca²⁺] curves with calculated EC₅₀ values. (e) Effect of 100 nM isoprenaline (Iso) on force at submaximal [Ca²⁺] (0.4 mM). Data are expressed as mean ± SEM. WT n = 15; KI-NT n = 8; KI-S282 n = 6; KI-D282 n = 7. **P < 0.01 vs. WT one-way ANOVA plus Dunnett’s multiple comparisons test (KI-NT, S282 and D282 vs WT EHTs). ^#P < 0.05 vs. corresponding baseline, paired Student’s t-test (panel e). Concentration response curves were fitted to the data points and curve comparison was done by using extra sum-of-squares F-test, log-EC₅₀ was different for each data set. P = 0.0008 (panel d).