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. 2021 Feb 19;12:1159. doi: 10.1038/s41467-021-21372-4

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© The Author(s) 2021, corrected publication 2021

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. 2 — a Optical cross-sections through the caudal PNP (at 75% of rostrocaudal PNP length, shown in schematic) of No Cre and Cre;Fl/− embryos at E9.5, visualised using reflection imaging. The white arrow indicates the quantified eversion of the apical neuroepithelium, which should elevate its lateral margins to form neural folds. Scale = 50 µm. b, c Quantification of neural fold elevation (b, positive values) or eversion (b, negative values) and mid-PNP neuroepithelial thickness (c) in control, Cre;Fl/Fl and Cre;Fl/− embryos at 16–20 somite stages, before the PNP is significantly longer in Cre;Fl/− embryos. The inset shows an optical cross-section through a phalloidin-stained PNP indicating the thickness of the neuroepithelium (cyan bracket). b Control = 7, Cre;Fl/Fl = 6, Cre;Fl/− = 7 embryos. c Control = 9, Cre;Fl/Fl = 6, Cre;Fl/− = 7 embryos. Scale = 50 µm. d Representative (>6 embryos) surface-subtracted images of the apical neuroepithelium in a Cre;Fl/Fl embryo and Cre;Fl/− littermate showing the apical marker ZO1 and recombined cells lineage traced with EGFP. Scale = 20 µm. e Quantification of the orientation of the apical long axis of neuroepithelial cells in No Cre, Cre;Fl/Fl and Cre;Fl/− embryos. Blue arrows indicate the median orientation; p values from Chi² with Yates continuity correction; EGFP and tdTom not significantly different within genotypes. No Cre 5592 cells from six embryos; Cre;Fl/Fl EGFP/tdTom 318/2373 cells from five embryos; Cre;Fl/− EGFP/tdTom 267/3799 cells from six embryos. RC rostrocaudal, ML mediolateral. f Apical surfaces from d colour coded to illustrate orientation. Bright colours indicate medio-lateral, dark colours indicate rostro-caudal orientation. Scale = 50 µm. g Quantification of neuroepithelial apical area. Each point represents the median value for an embryo and lines link points from the same embryo, n = 6 embryos per genotype. p values from ANOVA with post-hoc Bonferroni. Embryos compared had 15–20 somites and had been administered tamoxifen 24–28 h prior to analysis.