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. 2023 Feb 24;30(5):1155–1165. doi: 10.1038/s41418-023-01126-z

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© The Author(s) 2023, corrected publication 2023

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 Schematic representation of the treatment of murine tumor organoids seeded as single cells with recombinant TGFβ1 and timing of quantification of the morphology and gene expression signatures. B Representative brightfield images of YFP + EpCAM + (N = 6), YFP + EpCAM- (N = 6) and YFP + secondary (Met, N = 6) tumor organoids on day 3 post TGFβ1 treatment. The pre-treatment (PRE) organoid morphology is indicated. Representative images of organoids post-treatment (POST) are shown. Arrows indicate a branching phenotype. Images are representative of 3 biological replicates. Scale bar = 300 μm. C Quantification of the tumor organoid morphology 3 days post TGFβ1 treatment. YFP + EpCAM + (N = 6), YFP + EpCAM- (N = 6) and YFP + secondary (Met; N = 6) tumor organoids. Organoids are grouped as glandular (G) and solid (S) morphologies pre-treatment (PRE). Quantification of glandular (green), solid (red) and branching (blue) post cytokine treatment (POST). Data includes 3 biological replicates and is presented + /− SEM. ****p < 0.0001, Chi-square test. D, E mRNA expression levels of mesenchymal markers, Vim, Cdh2, Fn1, Snai1, Snai2, Zeb1 (D); and S100 family members, S100a4, S100a14 (E), in YFP + EpCAM + (E+), YFP + EpCAM- (E-) primary tumor organoids and YFP + secondary tumor organoids following the addition of TGFβ1 (blue). Each dot represents an individual organoid line. Organoids are grouped as glandular (G) and solid (S) morphologies. Data includes three biological replicates and is presented as log10 fold change relative to the vehicle (white) control, mean + /− SEM. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, paired t-test.