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. 2019 Jul 1;9:9475. doi: 10.1038/s41598-019-45851-3

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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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The lht mutant embryo exhibits defects in vascular maturation. Whole-mount immunohistochemistry for ZO-1 (a tight junction protein) at 2.2 dpf for WT and lht mutant embryos (A). Images are 3D projections of z-stack images acquired by confocal microscopy. Inset images display lumen formation in a blood vessel. Yellow circle depicts the vascular lumen in WT embryos (n > 10). In situ hybridization for EGFL7 expression in WT and lht mutant embryos at 2.3 dpf (n > 10) (B). Transmission electron micrograph of a transverse section of the dorsal aorta of WT and lht mutant embryos at 3.1 dpf (C; n = 2). In situ hybridization for etgase at 2.3 dpf (D). White arrows pointing at the CCV in WT represent the wide vascular lumen surrounded by etgase-positive cells. The white arrowheads in lht mutant embryos show disoriented etgase-positive cells around a putative blood vessel (n > 10). White dotted lines in the right-most panel indicate the vascular lumen. AA, aortic arch; LDA, lateral dorsal aorta; etgase, embryonic transglutaminase. Scale bars: 20 μm (A), 100 μm (B), 250 μm (C), 10 μm (B).