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. 2021 Feb 26;12:1328. doi: 10.1038/s41467-021-21557-x

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© The Author(s) 2021

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. 1 — a Collection of blastocysts after crossing Prdm14^mut/+ rats with Prdm14^HV/+ rats. Prdm14-H2BVenus positive blastocysts were selectively used for subsequent blastocyst injection. b Schematics showing injection of male (XY) or female (XX) Rosa26^{tdTomato/tdTomato} rat ESCs into Prdm14-H2BVenus positive blastocysts. c Chimerism of coat color, testis, and ovary in Prdm14^HV/+ chimeric rats with Rosa26^{tdTomato/tdTomato} rat ESCs. Scale bar in testis: 5 mm, in the ovary: 1 mm. d Immunohistochemical staining of testis and ovary of chimeras in Fig.1c. Some (filled arrowheads) but not all DDX4 positive germ cells originate from tdTomato positive donor ESCs. Scale bars: 100 μm. e Chimerism of coat color, testis, and ovary in Prdm14^mut/HV chimeric rats with Rosa26^{tdTomato/tdTomato} rat ESCs. Scale bar in testis: 5 mm, in the ovary: 1 mm. f Immunohistochemical staining of testis and ovary of chimeras in Fig.1e. All the DDX4 positive germ cells originate from tdTomato positive donor ESCs. Scale bars: 100 μm. g Offspring obtained by crossing Prdm14^HV/+ chimeric rats with wild-type Crlj: WI rats. Ubiquitous tdTomato expression shows successful germline transmission of injected Rosa26^{tdTomato/tdTomato} rat ESCs. h. Offspring obtained by crossing Prdm14^mut/HV chimeric rats with wild-type Crlj: WI rats. All the neonates ubiquitously expressed tdTomato indicating perfect germline transmission of Rosa26^{tdTomato/tdTomato} rat ESCs.