Fig. 4

Cxcr4 marks SHF progenitors in mouse PSC-derived spheroids and in embryos. a RNA-seq analysis of differentially expressed surface receptors between Hcn4-GFP⁺ and Tbx1-cre, RFP⁺ CPCs in vitro. We identified 240 differentially expressed surface receptors of which the top 55 are shown here (all with Benjamini–Hochberg adjusted p-value < 0.05). b qPCR analysis of Cxcr4 expression in Hcn4-GFP + and Tbx1-cre, RFP + CPCs. Data are mean ± SEM; n = 3; *p < 0.05; p values were determined using a paired Student’s t test. c Representative flow histograms of Cxcr4 staining of Hcn4-GFP⁺ and Tbx1-cre, RFP⁺ CPCs compared to unstained GFP⁺/RFP⁺ CPCs. d Representative flow cytometric analysis and sorting strategy of Isl1-cre, RFP⁺ CPCs (left) and Cxcr4^−/+ CPCs (right). e Clonal cell-fate assay showing qPCR results from 2 × 24 single-cell Cxcr4⁻ and Cxcr4⁺ clones, sorted and plated at day 5.5 and analyzed 7 days later. Solid colors represent expression of the gene of interest (Ct value < 30). Green: cTnT, Red: SM22, Blue: Pecam1, Yellow: FSP1/S100A4. f Cxcr4⁻/+CPCs isolated day 5.5, treated with EdU 24 h after isolation and stained for EdU and DAPI. Scale bars represent 50 μm, bar graph shows quantification of EdU + CPCs, Data are mean ± SEM; n = 4; *p < 0.05; p values were determined using a paired Student’s t test. g Microarray analysis showing expression of cardiac genes in Isl1-cre, RFP⁺ vs. Isl1-cre, RFP^- CPCs isolated day 5.5