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. 2021 Sep 29;11:19369. doi: 10.1038/s41598-021-98766-3

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© The Author(s) 2021

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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Changes in 3D HTM spheroid size (A) during their 3D culture in the presence or absence of DEX or TGFβ2, and representative 3D spheroid images stained by DAPI and phalloidin (B, C). Among the experimental groups treated without (CONT) or with 250 nM DEX or 5 ng/ml TGFβ2, changes in the mean size of 3D HTM spheroids during 3D culture at Days1, 3 and 6 were plotted in panel A. Representative immunolabeling XY axes plane image (panel B) and 3D XYZ axes image (panel C) of the 3D HTM spheroid (CONT) at Day 6 stained by DAPI and phalloidin. These experiments were performed in triplicate using fresh preparations (n = 10 or 5 for size measurement and immunolabeling, respectively). Data are presented as the arithmetic mean ± standard error of the mean (SEM). **P < 0.01; ***P < 0.005 (ANOVA followed by a Tukey’s multiple comparison test). Scale bar: 100 µm.