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. 2019 Jun 5;10(6):438. doi: 10.1038/s41419-019-1683-1

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© The Author(s) 2019

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. 3 — a Diagram illustrating the procedure of tetraploid aggregation, in which one Rbpj^−/− morula aggregated with two wild-type tetraploid EGFP-expressing four-cell embryos. The tetraploid cells contribute exclusively to the trophoblast cells of the placenta, rather than the allantois. b E10.5 embryos and placentas generated by tetraploid aggregation assay. c HE staining of E10.5 reconstructed placentas from tetraploid aggregation assay. d Shallow invagination of allantoic blood vessels into the chorionic ectoderm was revealed by HE and laminin staining in placentas with allantois-specific deletion of Rbpj by Sox2^cre/+. Cy3-labeled Laminin is in red, DAPI-labeled nuclei in blue. e The differentiation of a chorionic trophoblast was revealed by the expressions of Gcm1 and Synb (markers for SynT-II), and Syna (marker for SynT-I). By in situ hybridization, the expression of these markers was comparable between the Rbpj^f/f and Rbpj^f/f/Sox2^cre/+ placentas. Images in b–e are representative of at least three independent experiments. Al allantois, Cp chorion plate, Lab labyrinth, f fetal blood vessel, m maternal blood sinus, p placenta, Sp spongiotrophoblast layer. Yellow scale bars: 1 mm; white and black scale bars: 100 μm