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. 2022 Aug 15;21(11):1314–1323. doi: 10.1038/s41563-022-01323-0

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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. 4 — a, Schematic showing the regions measured by AFM, black arrows indicate the recorded region. b, Spread of data for each condition; red lines represent mean and whiskers s.d. (two-tailed t-test ****P < 0.0001, ***P < 0.0006, CI = 95%, n_{control mesoderm} = 8, n_{ca-Mypt1 mesoderm} = 7, n_{migratory NC in control} = 8, n_{migratory NC in ca-Mypt1} = 8 embryos; 64 indentations per embryo). c, Immunofluorescence analysis showing acetylated α-Tubulin and α-Tubulin signal in NC platted on soft or stiff hydrogels (nuclei in magenta and NC border in cyan) (scale bar, 20 μm). d, Normalized fluorescence intensity ratio of acetylated α-Tubulin versus α-Tubulin; red lines represent mean and whiskers s.d. (two-tailed t-test, ****P < 0.0001, n_soft = 13 clusters; n_stiff = 15 clusters). e–g, Piezo1 regulates microtubule acetylation in vivo. e, Schematic showing the plane of sectioning (HM, head mesoderm; ML, mediolateral; AP, anteroposterior; DV, dorso-ventral). f, Upper panel, confocal projections of transverse cryosections showing highlighted NC nuclei (cyan) and fibronectin (magenta); lower panel, colour-code image of the acetylated α-Tubulin channel (scale bar, 100 μm); upper right inset emphasizes the signal differences between both treatments in the NC (scale bar, 50 μm). g, Normalized acetylated (norm. acet.) α-Tubulin fluorescence intensity, spread of data is shown; red lines represent mean and whiskers s.d. (two-tailed t-test ***P = 0.004, CI = 95%, n_Control = 12, n_Piezo1-MO = 12 embryos). c and f are representative confocal projections from three independent experiments.

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