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. 2017 Nov 9;7:15136. doi: 10.1038/s41598-017-15427-0

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© The Author(s) 2017

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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Development of E3.5 ICMs. (A) ICM immediately after IS, and (B) 24 hrs after IS. Blue: Oct4, green: Gata4, white: nuclei; right panel shows merged pictures, (C) Time-lapse imaging of E3.5 ICM immediately after isolation, after 12 hrs and after 24 hrs of in vitro culture (single optical sections), (D) The scheme of the ultimate PE origin: (a) Pdgfrα ^H2B-GFP-expressing cells localised on the surface of isolated ICM from the beginning of in vitro culture, (b) Pdgfrα ^H2B-GFP-expressing cells which translocated from inside to outside, (c) Pdgfrα^H2B-GFP-negative cells which up-regulated Pdgfrα ^H2B-GFP during in vitro culture and migrated outside, (d) Pdgfrα^H2B-GFP-negative cells which up-regulated Pdgfrα on the surface of ICM and maintained this position, (E) The scheme of the fate of all Pdgfrα ^H2B-GFP-expressing cells: (a’) cells which were localised on the surface of ICM, contributing to PE, (b’) cells which were initially placed inside and translocated outside, contributing to PE, (c’) cells localised inside, which down-regulated Pdgfrα ^H2B-GFP during culture, (d’) cells which underwent apoptosis. Scale: 20 μm.