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. 2019 Dec 17;10:5754. doi: 10.1038/s41467-019-13623-2

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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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Fig. 2 — a Representative immunostaining for NCX1, RyR2, and LTCC (DAPI counterstain, blue) in hearts of 10-month-old mice. Scale bar, 100 μm. b Representative immunofluorescence (top: phalloidin, red; DAPI, blue (scale bar, 100 μm)) and electron microscopy (scale bar, 5 μm (middle) and 1 μm (lower)) images of myofilaments. c Frequency-dependent shortening of cardiomyocytes (n = 16 and 20 cells measured from 4 and 5 hearts, respectively). #P < 0.05 vs. floxed mice based on Student’s t-tests. (d) Indo-1 fluorescence in single cardiomyocytes stimulated at 1 Hz (1.8 mM Ca²⁺, n = 20 and 37 cells from 3 and 4 hearts, respectively; 5 mM Ca²⁺, n = 20 cells from 3 hearts per group). Normalized peak amplitudes (e), decay time constants (obtained by fitting to the decline phase) (f), and times to peak (g) of Ca²⁺ transients. h Estimation of sarcoplasmic reticulum (SR) Ca²⁺ content (n = 15 and 10 cells from 3 and 2 hearts, respectively). ^#P < 0.05 between indicated groups based on Student’s t-tests.