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. 2017 Nov 28;7:16508. doi: 10.1038/s41598-017-16642-5

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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 lethal mutant embryos and larvae grafted into rag1 mutants. (a,c) Morphology of 72 hpf spns2 mutant embryos (a) and 7 dpf oep mutant larvae (c) before and after transplantation into germ cell-depleted rag1 mutants. The spns2 mutant has cardia bifida (two hearts) as determined by cmlc2-driven mRFP expression (arrowheads), and the oep mutant exhibits a single eye (arrows). Scale bar: 1 mm. (b,d) Histology of organs found in grafted spns2 mutant embryos (b) and oep mutant larvae (d) at 2 months after transplantation. Arrowheads indicate each organ as labelled above the panels. Note that testes of the grafted spns2 and oep mutants contained all stages of spermatogenic cells: spermatogonia (g), spermatocytes (c), and sperm (s). The heart was not observed in grafted spns2 mutant embryos (b). Scale bar: 200 µm. (e) RT-PCR analysis of grafted spns2 and oep mutants at 2 months post-transplantation. Wild-type zebrafish at 2 mpf were used as a positive control. Negative controls lacking reverse transcriptase are in alternating lanes (−). Each experiment was conducted using two independent samples.