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. 2021 Feb 11;11:3588. doi: 10.1038/s41598-021-82954-2

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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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Disruption of the Smad4 gene in Sfrp5-expressing cells leads to hypoplastic heart formation and impaired cardiac differentiation. (A,B) Gross appearance (A) and Hematoxylin and Eosin staining (B) of sagittal sections of Smad4 mutant (Sfrp5 ^Cre/+; Smad4 ^del/del) and littermate control (Sfrp5 ^Cre/+; Smad4^del/+) embryos dissected at E9.5. Scale bar = 100 μm. (C) Relative gene expression levels of key factors involved in cardiogenesis. n = 9 and 7 for control and Smad4-cKO, respectively. Data are presented as mean ± SEM. (D) Immunofluorescent staining for cTnT (green) in control and Smad4 mutant embryos at E9.5. Samples were counterstained with 4′, 6-diamidino-2-phenylindole (DAPI) to visualize nuclei (blue). Scale bar = 50 μm.