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. 2021 Jan 27;155(5):581–591. doi: 10.1007/s00418-020-01960-z

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© The Author(s) 2021

Open AccessThis 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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Fig. 1 — Ishikawa spheroids in cell culture and morphological confirmation of cell polarity. a–c show the development of Ishikawa spheroids in cell culture from day 0 to day 6. Microscopic phase contrast images were inverted. Scale bars: 150 µm. d shows a representative polarized Ishikawa spheroid on day 6. Nuclei are stained with Hoechst 33,342 (blue), tight junctions with anti-ZO-1 antibody (green). Scale bar: 30 μm