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. 2019 Jul 24;9:10716. doi: 10.1038/s41598-019-47009-7

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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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Introduction of multiple expression cassettes in the same heart. (A) Diagram of plasmids used in cotransfection studies. (B) Single channel and merged image of palmTagRFP and palmEGFP expression 160 hrs. post transfection. (C) Flow cytometry plot of a dissociated heart 160 hrs. post transfection with the constructs from (A). (D) Detection of palmTagRFP, PalmEGFP, and copositive cells following dissociation and culture of transfected hearts. Scalebar = 50 um. (E) Diagram of a dual promoter construct. (F) Single channel and merged images of a heart 160 hrs. post transfection. (G) Flow cytometry plot of a dissociated heart 160 hrs. post transfection with the construct from (E). (H) Dissociation and culture of cells from a heart transfected with the dual promoter construct demonstrating the absolute intensity of turboRFP to EGFP intensity can vary among individual cells. Scalebar = 50 um.