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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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Lack of nuclear localization of Nkx2-5 in Smad4-cKO embryonic hearts. (A) Whole-mount ISH of control and Smad4-cKO embryos with a probe for Nkx2-5 at E9.5. (B) Immunofluorescent staining of sagittal sections of control and Smad4-cKO embryos at E9.5 using anti-Nkx2-5 antibodies (red) and DAPI for detection of nuclei (blue). Arrows indicate the lack of nuclear localization of Nkx2-5. H, heart. Scale bar = 100 μm. (C and D) Percentage of Nkx2-5 protein distributing cell in control and Smad4-cKO embryonic hearts (C) and the percentage of cells showing nuclear localization of Nkx2-5 in control and Smad4-cKO embryonic hearts (D). Data were collected from 10 sections for each embryo (n = 3 and 3 for control and Smad4-cKO, respectively). Data are presented as mean ± SEM, **P < 0.01 (Mann–Whitney U test).