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. 2020 May 12;11:2366. doi: 10.1038/s41467-020-16247-z

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© The Author(s) 2020

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. 4 — a, d Transverse sections of SG placode explant culture with DMSO (vehicle), Cytochalasin D, CK666, ethanol (vehicle), cyclopamine, or SU5402, as indicated (green: phalloidin; blue: DAPI) showing inhibition of invagination. Scale bars: 50 μm. b, c, e, f Quantification of placode depth (b, e) and protrusion abundance (c, f) in inhibitor-treated placodes vs. vehicle-treated controls (means ± SDs, n = 12 placodes, 4 placodes per treatment per experiment from 3 independent litters; n = 100 cells, ~8 per placode). c, f Quantification of by inhibitors compared to controls, g top-view live images of SG placodes in mT/mG explants treated with vehicle, cyclopamine or SU5402. Dashed line circles: placode region. Asterisks inside cells: the cell bodies of protrusive cells. Arrowheads: direction of the protrusion. “+”: the centre of the placode. Note that protrusions are centripetal in control and cyclopamine treated tissues, but not in SU5402-treated tissue. Only isolatedly labelled cells were analysed. Scale bars: 20 μm. h Orientation of protrusions in the three treatments (0° indicates centripetal) (data from three independent litters, six placodes in each condition). p Values in b, c, e, f are from two-tailed unequal variance t tests, in (h) are Rayleigh test. Source data are provided as a Source Data file.