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. 2022 Jun 22;12:10563. doi: 10.1038/s41598-022-14875-7

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© The Author(s) 2022

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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Identification of different cell structures in 3D chicken intestinal organoid and the monolayers. Immunohistochemistry images of 3D and 2D structures of chicken intestinal organoids, processed 2 days after seeding. (a,b) Progenitor cells are visualized with anti-Sox9. (c,d) Enteroendocrine cells are visualized by Chromogranin A. (e–g) Tight junctions are visualized with beta-catenin or occludin (h) Mucus-containing goblet cells are visualized using Periodic Acid Schiff (PAS)-positive cell staining of 2D chicken intestinal organoids. The actin filaments are visualized with rhodamine phalloidin (red), and nuclei are stained with DAPI (blue) Scale bar: 400 µm.