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. 2018 Mar 14;9(3):399. doi: 10.1038/s41419-018-0442-z

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© The Author(s) 2018

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. 1 — a Confocal images of GFP expression plus whole-mount and sectioned in situ hybridisation (ISH) of Bmp2 in E9.5 WT and Nkx2.5^Cre/+;Bmp2^tg/+ hearts. White arrowheads mark normal Bmp2 expression in AVC myocardium; black arrowheads mark ectopic Bmp2 expression in chamber myocardium of the transgenic heart. b Top two rows, hematoxylin and eosin (H&E) staining of general and detailed views (insets) of transverse and parasagittal sections of E10.5 WT and Nkx2.5^Cre/+;Bmp2^tg/+ hearts, showing mesenchymal cells in the AVC (black arrowheads) and also in the left ventricles of transgenic hearts (white arrowheads). Third row, whole-mount images of E14.5 WT and Nkx2.5^Cre/+;Bmp2^tg/+ embryos showing oedema in the dorsal region of the transgenic embryo (white arrowhead). H&E stained transverse sections show general and detailed views of ventricular and valve dysmorphology and a ventricular septal defect in the E14.5 Nkx2.5^Cre/+;Bmp2^tg/+ heart (asterisk). Fourth row, details of the atrioventricular (AV) valves (arrowheads) and left ventricle in the WT and transgenic heart; trabeculae (white arrowheads) are larger in the transgenic heart. The arrows mark the WT coronary vessels and the dysmorphic ones in the transgenic heart. c ISH showing Bmp2 mRNA in the AV valves in both E14.5 genotypes (arrowheads) and ectopically throughout the myocardium of Nkx2.5^Cre/+;Bmp2^tg/+ ventricles. d Staining for pSmad 1/5 (red), MF 20 (green), isolectin B4 (IB4, white) and DAPI (blue) in E14.5 hearts. General views of transverse heart sections. Note the ventricular septal defect (asterisk) in the transgenic heart. Detailed views show the right ventricle and AV valves of both genotypes, with discrete pSmad 1/5 staining in WT trabecular endocardium (white arrowheads), capillaries in the compact myocardium (yellow arrowheads) and AV valves mesenchyme (arrows, mesenchyme marked with a yellow asterisk). The Nkx2.5^Cre/+;Bmp2^tg/+ heart shows widespread pSmad1/5 expression both in ventricles (including cardiomyocytes, arrows) and in AV valves mesenchyme. e pSmad1/5 activation index in the trabecular and compact myocardium (Tm, Cm) of E14.5 WT and transgenic hearts. a atrium, AVC atrioventricular canal, IVS interventricular septum, la left atrium, lv left ventricle, mv mitral valve, ra right atrium, rv right ventricle, tv tricuspid valve, v ventricle; scale bars 200 μm