| Summary: | (A) (i-ii”) Immunodetection of GFP, the phosphorylated form of histone H3 (PH3) and DAPI staining on transverse section of chick embryo at 48hpe with the indicated plasmids. (iii) Quantification of the number of PH3 + cells in the GFP + cells in embryos expressing the indicated plasmids at 48hpe. (dots: embryo values; bar plots: mean ± s.e.m.). Data for pCIG , Pax3 , PAX3-FOXO1 samples are the same as in Fig 6Aiii . (B) (i-i”) Immunodetection of GFP, EdU and DAPI staining on transverse section of chick embryos 48hpe with the indicated plasmids and soaked with EDU 20h before harvest. (ii) Quantification of the number of EdU + cells in the GFP + population in embryos expressing the indicated plasmids (dots: embryo; bar plots: mean ± s.e.m.). (C) (i-i’) Z projection along the dorso-ventral axis of 3D scans of an embryos 72hpe with PAX3-FOXO1 and CCND1 . Dotted lines mark the apical cavity and the basal membrane (bm). (ii) Quantification of the distance of each GFP + nuclei and the apical surface at 72hpe with the indicated plasmids (Violin plots). (D) PITX2 detection via in situ hybridization on transverse sections of chick embryos 24hpe and 48hpe with the indicated plasmids and immuno-detection of GFP on the adjacent section slide. Bottom panels show areas on the embryos and section presented in the upper panels where PITX2 is expressed, including the lateral plate mesoderm (lpm) and the dorsal medial lip of the dermomyotome (dml). x’ and x” panels are blow-ups of a subset of x panel GFP + cells. Mann-Whitney U test p-value: **: p< 0.01, ****: p< 0.0001, ns: p> 0.05. Scale bars 50μm.
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