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. 2018 Apr 4;8:5632. doi: 10.1038/s41598-018-24014-w

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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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Rudhira is required for extra-embryonic vascular patterning. (a) Morphology of whole mount PECAM/CD31 stained yolk sac Red dots: branch points. Graph shows quantitation of branch points (N = 5 yolk sacs). (b) hematoxylin-eosin stained yolk sac sections. en: endoderm, m: mesoderm. (c,d) Placental morphology seen by histological analysis (c) or immunofluorescence staining (d) in sections of control and rudh^−/− as indicated. Boxed areas (in c) show magnified images of respective placenta. Immunostaining shows fetal and maternal blood cell pockets marked by Ly76. Nuclei are marked by DAPI (Blue). Arrows: maternal blood cells; arrowheads: fetal blood cells. de: decidua, sp: spongiotrophoblast, lb: labyrinth, ch: chorion. Graph shows quantitation of placental thickness. Results shown are a representative of at least three independent experiments with at least three biological replicates each. Statistical analysis was carried out using one-way ANOVA. Scale bar: (a,b) 200 µm; (c,d) 100 µm. *p < 0.05, **p < 0.01, ***p < 0.001.