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. 2019 Mar 14;9:4536. doi: 10.1038/s41598-019-40809-x

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© The Author(s) 2019

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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Cell type specific expression of Rspo3 and Wnt4. (a) Cell type specific expression of Wnt ligands. The violin and tSNE plots depict that Rspo3 mRNA expression is enriched in the fibroblast cells and Wnt4 mRNA expression is enriched in the epithelial cells. The expression level of violin plot is the natural log of the normalized UMI counts. *Bonferroni adjusted p value < 0.001. (b) RNA In situ hybridization confirms cell type specific expression of Rspo3 and Wnt4 in the organoid. The organoid FFPE section is hybridized and amplified by the probes specifically targeting Rspo3 and Wnt4 mRNAs. Rspo3 probes are the blue dots detected in the fibroblast cells surrounding the organoid, and Wnt4 probes are the red dots detected in the epithelial cells forming the wall of the organoid sphere (Scale bar = 100uM).