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. 2019 Dec 2;9:18130. doi: 10.1038/s41598-019-54544-w

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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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(a) Whole-mount in situ hybridization showing the expression of PGC marker genes, including nanos1 and vasa, after morpholino injection at 25 hpf. The reduction of nanos1 and vasa expression intensity was obviously observed in MO^aug-injected embryos. The nanos1 expression was also decreased in MO^sp-injected embryos and it was recovered when mRNA was co-injected. The expression levels of nanos1 (b) and vasa (c) were graded as the normal and abnormal percentage. The mean pixel intensity of nanos1 and vasa gene expression of the normal embryos were 24.7 ± 8.7 and 22.6 ± 8.2, respectively. An abnormal level of the gene expression was taken as one with the value less than the mean minus one standard deviation of the gene expression of the wild-type, i.e., less than 16 for nanos1 and 14.4 for vasa gene. The number of animals quantified in each group are shown on top of the bars.