Fig. 3.

Comparison of Isl1⁺ and Nkx2-5⁺ cardiac progenitor cells. a Confocal images showing nuclear-, cytoplasmic- and co-localization of GFP in CPCs FACS-sorted from Isl1^+/nGFP/Nkx2-5-emGFP⁺ embryos. Nuclei were stained with DAPI (blue). b Immunofluorescence-based quantification of (a). Isl1⁺Nkx2-5⁻, Isl1⁺Nkx2-5⁺ and Isl1⁻Nkx2-5⁺ cells were FACS-sorted from Isl1^+/nGFP/Nkx2-5-emGFP⁺ embryos at E8.5 and E9.5. Quantification of different cell populations was achieved by counting all immunostained cells in a multiwell dish. Mean ± s.d. are shown. Circles represent results from different biological replicates [n = 3; Σ (cell number) of E8.5 = 225, 260, 100; Σ (cell number) of E9.5 = 175, 180, 100]. c Clustering of Isl1 and Nkx2-5 co-expressing cells in Nkx2-5⁺ and Isl1⁺ CPC subpopulations. Cells that are not double-positive are labeled in gray, and clusters are indicated by colored circles. d, e Plots showing the predicted diffusion pseudotime of Nkx2-5⁺ cells projected on t-SNE plots of Isl1⁺ cells, and the expression of Isl1⁺ (d) and Nkx2-5⁺ (e). Expression levels of Isl1 and Nkx2-5 in CPCs are represented by a color spectrum as indicated. EC, endothelial cell. CM, cardiomyocyte