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. 2021 Jan 25;12:560. doi: 10.1038/s41467-021-20906-0

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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 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. 3 — a Top: the scheme for the Gata4 conditional knock out (Gata4 cKO) experiment in the embryonic stomach. Bottom: whole-mount X-gal staining on the stomach of the Gata4 cKO (E9.5) embryo at E18.5. Scale bar, 1 mm. b Fluorescent images from the Gata4-td Tomato of the control and Gata4 cKO (E9.5) stomachs at E18.5. Presented data are a representative image of n = 5 embryos out of three independent experiments. Scale bar, 1 mm. c IHC analyses of KRT7 for the stomachs of the Gata4 cKO (E9.5) embryo at E18.5. Arrows indicate the ectopic KRT7⁺ transitional epithelium in the distal stomach. Presented data are a representative image of n = 5 embryos out of three independent experiments. Scale bar, 500 µm. d IF analyses of KRT7, CLDN18, and GATA4 for the distal stomach of the Gata4 cKO (E9.5) embryo at E18.5. Presented data are a representative image of n = 5 embryos out of three independent experiments. Scale bars, 100 μm. e IF analyses of KRT7, KRT14, and GATA4 (top), or SOX2, P63, and GATA4 (bottom) for the stomach of the Gata4 cKO (E9.5) embryo at E18.5. Presented data are a representative image of n = 5 embryos out of three independent experiments. Scale bars, 100 μm. f Top: the scheme for the Gata4 overexpression (Gata4 OE) experiment in the embryonic stomach. Bottom: IHC analysis of CLDN18 for the Gata4 OE stomach at E18.5. Presented data are a representative image of n = 4 chimeric embryos out of three independent experiments. Scale bars, 500 μm. g IF analyses of SOX2, GATA4, and HA for the proximal stomach of the Gata4 OE embryo at E18.5. Presented data are a representative image of n = 4 chimeric embryos out of three independent experiments. Scale bars, 100 μm. h A scheme illustrates the Gata4-mediated specification of the primitive transitional epithelium into the columnar epithelium. TE transitional epithelium, CE columnar epithelium.