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. 2022 Aug 25;610(7930):143–153. doi: 10.1038/s41586-022-05246-3

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© The Author(s) 2022

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 — a,b, Sagittal sections of natural embryos at E8.5 (a) and day 8 ETiX embryoids (b) showing SOX2, SOX17 and GATA4. Scale bars, 100 μm. n = 3 ETiX8 from 3 experiments, n = 2 natural embryos. fg, foregut; hg, hindgut. c, Dot plot showing levels and proportion of cells expressing indicated genes in selected tissues of natural embryos and ETiX embryoids by inDrops scRNA-seq. d, UMAP of tiny-sci-RNA-seq dataset showing VE, gut and early development cell types. e, Day 6 ETiX embryoid (top) and natural E7.5 embryo (bottom) showing STELLA, NANOG and SOX2, highlighting the presence of committed PGCs (n = 9 ETiX6 from 2 experiments, n = 4 embryos). Outlined regions are magnified on the right. f, Quantification of PGCs during ETiX embryoid development (n = 9 ETiX6, 4 E7.5, 2 ETiX7, 4 E8.0, 4 ETiX8 and 3 E8.5). PGCs were scored for STELLA, NANOG and SOX2 expression. Violin plots show median and quartiles. Two-sided Mann–Whitney U-test, P = 0.3375 for ETiX6/E7.5 total STELLA-positive cells, P = 0.3042 for ETiX6/E7.5 triple-positive cells, P = 0.2277 for ETiX8/E8.5 total STELLA-positive cells, and P = 0.2536 for ETiX8/E8.5 triple-positive cells.

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