(A) Diagrams of the E8.5 (2–5 s) embryo showing the location of neuromesodermal and lateral/ventral-fated progenitors and the terminology used in this study. NSB, node-streak border; CLE, caudal …
Grafts of primitive streak and tail bud regions to the kidney capsule. (A) Masson’s trichrome-stained tumour sections derived from the indicated embryonic tissue regions. (B) Percentage of tumours …
(A) In situ hybridisation for Sox2 and T. Abbreviations are the same as in Figure 1D. Noto, notochord; tbm, tail bud mesoderm. (B–F) Confocal sections of wholemount, immunostained embryos; …
(A) Double in situ hybridisation for Sox2 and T at the end of axis elongation (E13.5, 65s). (B) Workflow for Sox2/T quantification and 3D reconstruction. (C and F) Total cells analysed per embryonic …
(A) 3D reconstruction of the E8.5 caudal region. (Aa–b) Dorsal view. (Ac) Frontal view along the PS and CLE. Colours show different thresholded Sox2/T populations. (B–C) Quantification of different …
(A–C) 3D analysis showing the relative levels of Sox2 and T protein in the E8.5–10.5 caudal region. Sox2+T+ cells express low-to-medium levels of both transcription factors (green cells in Ac, Ad, Bc…
(A) Representative examples of homotopic CLE grafts (numbers identify individual embryos in B–D). Cell fate in the rostral CLE in rostral-to-caudal (Aa–l) and medial-to-lateral direction (Am–t). …
Presomitic mesoderm contamination in CLE grafts.
Additional information and analysis to show the majority of paraxial mesoderm in CLE grafts is derived from NMPs rather than contaminating PSM progenitors that were co-grafted.
Graft donor tissue was examined for the presence of Sox2/T coexpressing cells. L1-3 and L/St5 pieces were dissected as described above. The underlying presomitic mesoderm was manually removed in the …
(A) Schematic diagram summarising the different grafting experiments conducted in this work and the embryos series they correspond to. Diagrams represent the primitive streak region at E8.5, with …
(A–C) Immunohistochemical confirmation of differentiation markers in different graft series. Upper row, DAPI-counterstained images (grey). Boxes, region magnified in lower row. Lower row, …
(A-B) Scoring of GFP+ cell contribution to the paraxial mesoderm in CLE homotopic grafts (embryo series 1 and 2). No PXM contribution was observed in L/St5 homotopic grafts. Graph format is the same …
(A) Representative examples of heterotopic CLE grafts (numbers identify individual embryos in B). Three different contribution patterns were observed: mainly neural (Aa-d), neuromesodermal (Ae–h) …
(A) Quantitative analysis of L1-3 graft contribution. Left, the percentage of neural (N) vs paraxial mesoderm (PXM) contribution in all scorable sections. Right, the percentage of embryos with …
(A) Representative examples of L1-3 to St1-3 grafts. (Aa and Af) Cells immediately after grafting. βcatCKO:sGFP grafted embryos were grown for 8 hr in the presence of 5µM 4-OHT after which the …
(A) Schematic diagram of mouse cross to obtain βcatCKO:sGFP embryos. Mice, homozygous for both a silent GFP (Gilchrist et al., 2003) and floxed β-catenin alleles (Brault et al., 2001) were crossed …
(A–C) Grafting scheme of L1-3 (A) or L/St5 (B) to St1-3 heterotopic grafts. Either AGFP7 control or βcatCKO:sGFP donor tissue was used to transplant into WT host embryos. (C) After receiving the …
(A) Boxplot for T intensity in notochord (noto), presomitic mesoderm (PSM) and T- (Neg) cells (n, number of nuclei sampled from wholemount stained embryos). The threshold for positivity was set at 7 …
(A) Representative examples of L/St5 heterotopic grafts (numbers in brackets identify individual embryos depicted in (B–C). (Aa, Af and Ak) Cells immediately after grafting. Arrowheads show grafted …
(A) Percentage of total embryos and sections containing GFP+ cells in the lateral/ventral mesoderm. LVM contribution is very rare in L1 grafts. Note that the L2–3 grafts produced LVM at different …
(A) Different L/St5 grafts performed. (B) Representative sections taken from grafts in A. Green, grafted cells; cyan, autofluorescence channel; arrows, notochord position; nt, neural tube; pxm, …
(A) NMPs and LPMPs coexist in the caudal progenitor zone. The NSB and L1 regions show NM fate. NM potency is broader, encompassing L2–3 regions and coincides with a Sox2+T+ phenotype. LPMPs reside …
3D reconstruction of the caudal progenitor region at different stages of axis elongation. (00:00) E8.5 (2-5s); (00:38) E9.5; (01:02) E10.5; (01:26) E11.5; (01:47) E12.5; (02:08) E13.5 (62-63s); …
3D reconstruction of the caudal progenitor region. (00:00) E9.5 WT sample and (00:28) example of a βcatCKO embryo at E9.5, after it was cultured in vitro for 24 hr in the presence of 5µM 4-OHT.