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. 2020 Aug 14;10:13810. doi: 10.1038/s41598-020-70655-1

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© The Author(s) 2020

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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Promotion of the development of colonic organoids by Tsc2 ablation. (a) Representative phase-contrast images of colonic organoids derived from the colon of 9- to 13-week-old control or Tsc2 CKO mice at 1, 3, 5, 7, and 9 days after cell plating. The white arrowheads indicate cryptlike buds. Scale bar, 100 μm. (b) Immunofluorescence analysis of Ki67 (red) and β-catenin (green) in colonic organoids cultured for 7 days as in (a). Scale bar, 100 μm. (c) The number of colonoids was determined as a percentage of colonic organoids at 3 days after cell plating as in (a). Data are means ± s.e. for a total of 105 (control) or 119 (Tsc2 CKO) organoids in three independent experiments. *P < 0.05 (Student’s t test). (d) The number of buds per colonic organoid was determined at 9 days after cell plating. Data are means ± s.e. for a total of 38 (control) or 28 (Tsc2 CKO) organoids in three independent experiments. ***P < 0.001 (Student’s t test).