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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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Growth factors RSPO1 and WNT3, from murine or chicken origin, are important for establishing intestinal organoid growth. (a) Immunoblot analysis of chicken RSPO1 and chicken WNT3 protein expression in transfected Hek293t cells. Actin was used as the protein loading control and anti-flag HRP was used to visualize RSPO1 and WNT3. Empty Vector (E.V.) is taken along as a control. (b) Brightfield images of mouse intestinal organoids (I,II) and chicken intestinal organoids (III,IV) at passage 2 (5 days after passage). Cultured in Matrigel with ICM, supplemented with Rspo1 and Wnt3 of murine origin, (c) or supplemented with chicken-derived RSPO1 and WNT3. Images and immunoblot are representative of three independent cultures. Scale bar 200 µm.