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. 2019 Aug 1;9:11191. doi: 10.1038/s41598-019-47665-9

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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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Wnt signaling is required for the differentiation of CNC into head cartilage structures. Snai2:eGFP or Wnt reporter embryos were treated with XAV939 (B,B’, F,F’, 20 μM; J,J’, 5 μM), IWR1-endo (C,C’, G,G’, 40 μM; K,K’, 10 μM) or DMSO (vehicle control) from stage ~28 to ~35. Embryos were washed and cultured again to the indicated stages, and images were taken with a Zeiss Axiozoom.V16 epifluorescence microscope. A representative embryo from each treatment group is shown on the left, with upper and lower panels displaying ventral and dorsal views (with anterior at the top), respectively, of the same embryos, and statistics is shown in the graphs on the right. ***P < 0.001.