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. 2019 Mar 5;9:3594. doi: 10.1038/s41598-019-40136-1

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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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Vegfaa and Vegfab are necessary for islet vessel development. (a) RT-PCR of vegfaa and vegfab on sorted 2 dpf, 3 dpf, and adult beta-cells. Full length gel is presented in Supplementary Fig. 3. (b) Phenotypic score of islet vessels in non-injected, control morpholino and vegfaa, vegfab, or vegfaa/vegfab morpholino injected Tg(fli1:EGFP; insa:tagRFP) embryos. Phenotypes are scored such that no phenotype is comparable to wildtype (more than 7 endothelial cells adjacent to beta-cells), mild phenotype (4–7 endothelial cells adjacent to beta-cells), and severe phenotype (less than 4 endothelial cells adjacent to beta-cells). (c) The number of beta-cells in 72 hpf Tg(fli1:EGFP; insa:tagRFP) control and morpholino injected embryos. In the vegfaa, vegfab, and vegfaa/ab morpholino injected embryos, only the embryos that demonstrated a reduction or absence of islet vessels were counted. n = 9–21. Box-and-whisker plots show median, and circles represent individual zebrafish. (d–h) Confocal projections of (d) non-injected, (e) scrambled injected, (f) vegfaa, (g) vegfab, or (h) vegfaa/ab morpholino injected embryos at 72 hpf; endothelial cells (green), beta-cells (red), and DAPI (grey). Scale bar = 10 μm.