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. 2019 Oct 10;9:14490. doi: 10.1038/s41598-019-50899-2

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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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Physiological analysis of hESCs-CMs treated with mBax_499 mRNA. Untreated cells and cells treated with mBax_499 mRNA for 24 h were examined for changes in calcium homeostasis and mechanical contraction. All slides were stimulated with 1 Hz and [Ca²⁺]_i transients and contraction signals were recorded and analyzed. (a) Representative illustration of average transients from untreated cells (black) or hESC-CMs treated with mBax_499 mRNA (gray). Left sided graph shows average of raw traces and the right graph shows super imposed traces (normalized to peak high). (b–e) Parameters measured from calcium transients: (b) Half width was calculated as duration from 50% time to [Ca²⁺]_i peak to 50% time to calcium decay. (c) Tau characterized the speed of calcium reuptake. (d) Basal [Ca²⁺]_i during relaxation. (e) Magnitude of transients represents the % of [Ca²⁺]_i alternations during contraction, calculate as ΔF/F₀ and normalized to untreated cells. (f–h) Parameters measured from contraction traces: (f) Representative illustration of average super imposed transients of mechanical contraction (normalized to peak high). (g) Half width was calculated as duration from 50% time to contraction peak to 50% time to relaxation. (h) Fractional shortening (%) characterized the percent change during contraction, normalized to untreated cells. All bar graphs indicate mean (SE). Data was collected from at least 5 different zones obtained from 5–6 independent slides. Parameters were calculated from an average of 15–20 successive transients for each zone. Two-tailed unpaired student’s t-test was performed to ensure no statistical differences.