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. 2020 Sep 7;11:4435. doi: 10.1038/s41467-020-18288-w

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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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Fig. 2 — a Fluorescent in situ hybridization (FISH) showing expression of integrin-alpha-6 (ia6, green) and histone 3 (h3, red) during stages 0, 1, 2, and 3 of WBR. White arrows indicate ia6+ cells beginning to cluster during stage 2. As clusters increase in size, they begin to lose ia6 expression. As cell clusters differentiate to form the blastula-like structure, ia6 expression is not detected (yellow arrows). DNA was stained with Hoechst (blue) in all panels. Blue dashed lines outline blood vessel boundaries. Images are representatives of four independent experiments. Scale bars 20 μm. b Single-positive (ia6, pou3, or h3) and double-positive (ia6/h3 or pou3/h3) cells were counted using the cell counter feature in FIJI, and for each stage, four images from four independent samples were counted. Graph shows percentages of ia6/h3 double-positive or pou3/h3 double-positive cells among all ia6+ or pou3+ cells. Source data are provided as a Source Data file.