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. 2023 Dec 20;625(7993):126–133. doi: 10.1038/s41586-023-06850-7

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© The Author(s) 2023

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Extended Data Fig. 5 — a. Time series of cytonemes in wild-type zebrafish embryos. Embryos were injected with Wnt5b-GFP/Ror2/mem-mCherry in the clone. At 6hpf, the embryos were treated with DMSO or JNK inhibitor SP600125 from 0’00” to 35’00” and imaged live. Yellow arrows indicate the retraction of cytonemes at the corresponding time points. b. The quantitative analysis of cytoneme length at the different time points in zebrafish embryos was treated as indicated in (a). (Wnt5b/Ror2: n = 25, 23, 16, 13; Wnt5b/Ror2 + SP600125: n = 35, 34, 38, 54, n = numbers of cytonemes in three biological repeats). Significance is calculated by two-way ANOVA together with Dunnett multiple comparisons test. p-values as indicated. c,d. PAC2 cells were transfected with indicated constructs and processed with a Wnt5a/b (c) and a Ror2 (d) antibody and phalloidin 405. Scale bar 20 µm. e. Fluorescence intensity of secondary antibody in IRSp53^4K-GFP and CDC42^T17N-GFP transfected PAC2 cells processed with Wnt5a/b antibody. (n = 28,23, 26, n = 28,31,22, n = number of cells in three biological repeats). Significance is calculated by the Kruskal-Wallis non-parametric test with a Bonferroni correction for multiple comparisons. f. Fluorescence intensity of secondary antibody in IRSp53^4K-GFP and CDC42^T17N-GFP transfected PAC2 cells processed with Ror2 antibody. (n = 23,26,23, n = 23,32,29, n = number of cells in three biological repeats). Significance is calculated by the Kruskal-Wallis non-parametric test with a Bonferroni correction for multiple comparisons. Box and whisker plots (b,e,f) show median, upper and lower quartile range with whiskers for minimum and maximum values.

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