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. 2022 May 12;13:2594. doi: 10.1038/s41467-022-30197-8

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© The Author(s) 2022

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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Fig. 9 — a, b Localization of EGFP-Toca1 in injured aISVs. a Confocal z-projection images of pre- and post-injured aISVs in the Tg(fli1a:EGFP-toca1);(fli1a:lifeact-mCherry) larva at 3 dpf and its subsequent time-lapse images at the indicated stages of repair. Lateral view, anterior to the left. Images of EGFP-Toca1 (green) and Lifeact-mCherry (red) fluorescence are merged. EGFP (top), mCherry (middle), and their merged (bottom) images corresponding to boxed areas are enlarged on the right. Arrow indicates blood flow direction. b Number of EGFP-Toca1 clusters colocalizing with Lifeact-mCherry-marked F-actin in vascular regions within 20 µm from the leading edge at approximately 25% reparative phase. Each dot represents an individual injured vessel (red and blue, upstream and downstream injured vessels, respectively). Data are means ± s.e.m. (n = 8 animals for each). *p < 0.05 by Welch’s two-sided t test. c, d Elongation of injured aISVs in toca1 mutant zebrafish. c Confocal z-projection images of injured aISVs and those after 6 h in wild-type (left), toca1^nf4/+ (middle), and toca1^nf4/nf4 (right) larval zebrafish at 3 dpf with Tg(fli1a:lifeact-mCherry) background are as in a. Red and blue arrowheads, leading edges of upstream and downstream injured vessels, respectively; green arrowhead, anastomotic site of the injured vessel. d Quantification of elongation lengths of upstream and downstream injured aISVs in wild type, toca1^nf4/+, and toca1^nf4/nf4 embryos, as in c. Each dot represents an individual larva (red and blue, upstream and downstream injured vessels, respectively). Data are means ± s.e.m. (wild type, n = 13; toca1^nf4/+, n = 25; toca1^nf4/nf4, n = 16). *p < 0.05, **p < 0.01 by one-way ANOVA followed by Tukey’s test among the upstream or downstream group. Source data are provided as a Source data file. Scale bars: 10 µm (a, c), 5 µm (enlarged images in a).