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. 2021 Aug 26;12:5136. doi: 10.1038/s41467-021-25370-4

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© The Author(s) 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 Confocal maximum intensity projection images of E8.5 tailbuds probed with T-BRA (green), SOX2 (red) and TBX6 (magenta) antibodies and counterstained with DAPI (grey) reveal the enlarged CLE and accumulation of T-BRA^pos, SOX2^pos, TBX6^pos cells in Pbx-com and Pbx-DKO tailbuds. The cartoons on the right depict dorsal views of E8.5 control and Pbx mutant tailbuds, with NMPs represented as red dots and MPCs as blue dots. Scale bars: 50 µm. b Confocal transverse sections of control, Pbx-com and Pbx-DKO tailbuds through the caudal progenitor zone. Black dashed lines in the cartoon indicate the plane of sections. Representative sections S1 and S2 reveal accumulation of T-BRA/SOX2 double-positive (T-BRA^pos;SOX2^pos) NMPs in Pbx mutants (yellow arrowheads). TBX6 staining shows reduced expression in PSM and co-localisation with NMP markers in Pbx-com and Pbx-DKO tailbuds (white arrowheads). The panel on the left summarises the molecular signature used to identify the different cell types along the path towards PSM. Scale bars: 30 µm. c Box plots assessing the proportion of T-BRA^pos;TBX6^pos cells (MPCs) relative to DAPI^pos cells (total cells) or to TBX6^pos cells (PSM), and T-BRA^pos;SOX2^pos cells (NMPs) and TBX6^pos cells (PSM) relative to DAPI^pos cells. The box contains the 25th to 75th percentiles of the dataset, the centre line denotes the median value (50th percentile) and the whiskers extend from the smallest to the largest value. P-values were calculated by one-way ANOVA and Tukey’s test for multiple comparisons and are indicated in the figure. d Distribution of NMPs and MPCs along the anterior/posterior axis shows accumulation of NMPs and MPCs in the posterior tailbud of Pbx-com and Pbx-DKO embryos. Data are mean ± s.e.m. Black dashed lines in the cartoon indicate the plane of sections. In c and d, the percentage of different cell types for each embryo was calculated from 10 independent sections equally distributed along the anterior/posterior axis. In a, b and d: CLE caudal lateral epiblast, NSB node-streak border, PS primitive streak, PSM pre-somitic mesoderm, Som somites, NT neural tube, A anterior, P posterior, D dorsal, V ventral.