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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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Failure of organ formation in grafted blastular and gastrular embryos. (a,b) Representative 1000-cell-stage (3 hpf) embryos (a), shield-stage (6 hpf) embryos (b) and grafted embryos two months after transplantation. Both embryo stages were subcutaneously transplanted into rag1 mutants. Scale bar: 1 mm. (c,d) Histology of 1000-cell-stage embryos (c) and shield-stage embryos (d) before and after transplantation. Sections were stained with haematoxylin and eosin. Although brain-like organs with an irregular shape and cell body arrangement were observed in all of the grafted early-stage embryos, other organs were not observed, except an eye (arrow) and intestine (arrow head) in a few grafted embryos. Scale bars: 200 µm (brain-like organ and eye), 100 µm (intestine), 50 µm (blastula and gastrula). (e) RT-PCR analysis of organ-specific genes in the grafted blastular and gastrular embryos at 2 months post-transplantation. Each experiment was conducted using two independent samples. Wild-type zebrafish at 2 mpf were used as positive controls. Negative controls lacking reverse transcriptase are located in alternating lanes (−). gapdh was used as a positive control.