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. 2021 Feb 24;11:4430. doi: 10.1038/s41598-021-83398-4

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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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Co-culture of cardiomyocytes and ChR2-transduced myofibroblasts. (A–C) Confocal image of co-culture of MFBs and CMs. α-actinin (A, red) marks CMs, and α-smooth muscle actin (B, α-SMA, green) marks MFBs. (C) Merge of (A) and (B) with DAPI (blue) to stain nuclei. (D) Phase-contrast image of CMs co-cultured with ChR2-MFBs. (E) Fluorescence image of same sample and area as in D, with YFP marking transduced MFBs. (F–H) Confocal images from 18 μm-thick z-stack of transduced MFBs and CMs. ChR2-YFP (green) marks transduced MFBs, α-actinin (red) marks CMs, and violet shows connexin43 (Cx43). ( F) CM layer of z-stack showing gap junctions between CMs. (G) Image from 4 μm above F showing MFBs on top of CMs, as well as Cx43 puncta, apparently between CMs and MFBs. (H) Maximum intensity projection of entire z-stack. Each layer of the z-stack is shown in Supplementary Fig. 1.