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. 2018 May 17;8:7753. doi: 10.1038/s41598-018-25964-x

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© The Author(s) 2018

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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POU5F1 expression is required for blastocyst formation in bovine embryos. (A) Immunofluorescence analysis of POU5F1 (green) and CDX2 (red) expression in D7.5 expanded blastocysts from the POU5F1 targeted knockout and injected control (IC) groups. (B) Genotype analyses from D7.5 blastocysts (n = 2) following confirmation of POU5F1 expression in embryos targeted for knockout. (C) Proportion of morulas collected on D7.5 that were stained and determined as POU5F1 positive (+/+, +/−) or negative (−/−). (D) Embryos in the targeted knockout group that failed to form blastocysts by D7.5 were genotyped to determine total mutation rate and allelic knockout efficiency (bi-alleic or monoallelic). (E) Immunofluorescence analyses of POU5F1 (green) and DAPI (blue) staining in D7.5 arrested morulas from targeted knockout and injected control groups. Scale bar = 100 µm.