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. 2018 May 10;37(33):4518–4533. doi: 10.1038/s41388-018-0295-z

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© The Author(s) 2018

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. 5 — MASTL overexpression increases proliferation and impairs organoid architecture in 3D culture. Cell lines were seeded into matrigel (Corning) and cultured under reduced serum and EGF conditions. a-c Phase contrast photos taken every day for 7 days and 20–40 individual colonies for each condition and time-point were measured in ImageJ for area and roundness. Shown are mean ± SEM from three independent biological replicates. For a, two-way ANOVA with multiple corrections, *p < 0.05, ****p < 0.0001. For c, unpaired t-test, *p < 0.05. d Seven-day acini were fixed and stained with H33342 (cyan) and phalloidin (red) and analysed by confocal microscopy with 0.3 µm z-stacks from the top of the acini to bottom shown, along with 3D projections and XY/YZ cross-sections to identify cleared lumens. e 3D acini from d were analysed using Imaris and individual nuclei were counted and measured for shape (ellipticity) and size (unpaired t-test, ***p < 0.001, ****p < 0.0001)