Fig. 6

Transcriptomic analyses. A–C PCA loading plot of the first two principal components distribution of EL (red), differentiated hPSC (blue) and undifferentiated hPSC (black) (A). Distribution of EL-ECs (mauve), EL-HSCs/HPs (purple), EL-Pre-HSCs (pink) and EL-mature HCs (red) (B). Distribution of hPSC-derived populations, hPSC-BCs (blue), hPSC-BC-CD144+ CD45+ (grey), hPSC-EB-CD144⁺ (orange) and hPSC-ECs (yellow) (C), each square represents one sorted cell population. D Visualizing significant differential level expression of selected genes of hPSC-BC-CD144⁺CD45⁺ and hPSC-ECs between hPSC-EB-CD144⁺ revealing distinct endothelial, hemogenic endothelial and hematopoietic signatures. All fold-changes were calculated with respect to the mean value of hPSC-EB-CD144⁺. P value ≤ 0.05. E–G GSEA curves for the most significantly enriched hematopoietic and endothelial GO biological process in hPSC-EB-CD144⁺ (E), hPSC-ECs (F) and hPSC-BC-CD144+ CD45⁺ (G) compared to undifferentiated hPSCs. Enrichment analysis was performed using Gene Set Enrichment Analysis software v4.1.0 and probing the c5.go.bp.v7.4 collection of the Molecular Signatures Database (MSigDB). GO = Gene Ontology; NES = normalized enrichment score. P value ≤ 0.05. H Non-supervised hierarchical clustering of transcript (rows) and samples (columns) based on their distance using Pearson correlation. Color intensity indicates expression levels scaled to the column mean. The heatmap is coded red for increasing and green for decreasing. The colored bar on top indicates how the heat map colors are related to the standard score (z-score), i.e., the deviation from row mean in units of standard deviations above or below the mean. I Model summarizing transcriptomic results where hPSC-EB-CD144⁺ give rise to hPSC-ECs, hPSC-BCs and hPSC-BC-CD144⁺CD45⁺, these two latter containing primitive and definitive hematopoietic progenitors, respectively