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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 essential for sprouting angiogenesis. (a) Migration tracks of control and rudh^−/− yolk sac endothelial cells subjected to wounding assay. Quantification showing the rate of migration and directionality compared between control and rudh^−/− yolk sac endothelial cells (marked by DiI-Acetylated LDL). Dotted line indicates the wound margin. Error bar indicates mean ± SD of a total of 14 cells. (b) NS and KD SVEC cells were tested for invasion using a Matrigel transwell-invasion assay. Graph indicates the relative invasion of cells 24 h post seeding as measured by Crystal Violet absorbance at 590 nm (c) NS or KD ECs were compared for adherence when plated on gelatin or collagen. Graphs indicate the relative adhesion of cells in 30 and 60 min post seeding as measured by Crystal Violet absorbance at 550 nm. (d) Collagen-based spheroid sprouting assay with spheroids formed from non-silencing (NS) and rudhira shRNA (KD) SVEC lines. Graph shows quantitation of the number of primary sprouts over time in days. Error bars indicate standard error of mean (SEM). Results shown are a representative of at least three independent experiments with at least three biological replicates. Statistical analysis was carried out using one-way ANOVA. Scale bar: (a,b,d) 100 μm. *p < 0.05, **p < 0.01, ***p < 0.001.