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. 2023 Dec 12;14:8245. doi: 10.1038/s41467-023-43999-1

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© The Author(s) 2023

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. 5 — a Representative calcium transient traces within d30 human heart organoids from each condition (n = 12 independent organoids per condition across three independent experiments). Traces represent data from an individual cardiomyocyte within human heart organoids. b Quantification of peak amplitude of calcium transient traces from each condition (n = 12 across three independent experiments). Values = mean ± s.e.m. c Quantification of calcium transient peak frequency for each condition (n = 12 independent organoids per condition across three independent experiments). Values = mean ± s.e.m. d Feature plots displaying key electrophysiological genes differentially expressed in the VCM and ACM clusters in each condition. e mRNA expression of key electrophysiological genes between d20 and d30 for each condition (n = 8 independent organoids per day per condition per gene across three independent experiments). Data presented as log₂ fold change normalized to d20. Values = mean ± s.e.m. f Representative voltage tracings of organoids in the EMM2/1 and control conditions depicting atrial-, nodal-, and ventricular-like action potentials (n = 8 (EMM2/1, Ventricular), 9 (EMM2/1, Atrial and Nodal; Control, Atrial), and 10 (Control, Nodal) individual cells from three independent organoids across three independent experiments). Representative immunofluorescence images (g) and quantification (h) of caveolin-3 puncta for each condition (n = 15 independent organoids per condition across three independent experiments). Green = caveolin-3, red = TNNT2, blue = DAPI. Scale bar = 20 μm. Data presented as fold change normalized to control. Values = mean ± s.e.m., one-way ANOVA with Dunnett’s multiple comparisons tests. Representative immunofluorescence images (i) and quantification (j) of KCNJ2+ puncta for each condition (n = 14 independent organoids per condition across three independent experiments). KCNJ2 = green, TNNT2 = red, DAPI = blue. Scale bar = 20 μm. Data presented as fold change normalized to control. Values = mean ± s.e.m., one-way ANOVA with Dunnett’s multiple comparisons tests. Source data are provided as a Source Data file.