Fig. 3: Differential responses of human arterial and venous endothelial cells to 1% oxygen.

a Schematic of the endothelial differentiation system. Human pluripotent stem cells were differentiated into primitive streak, then dorsal lateral mesoderm, and subsequently into arterial and venous endothelial cells²³. b Heatmap showing the expression profile of upregulated genes associated with angiogenesis, blood vessel morphogenesis and hypoxia in BCAS venous cells, and mapped onto the single-cell transcriptomics of human pluripotent stem cells, primitive streak, lateral mesoderm, pre-vein, and venous endothelial cell populations. The color intensity indicates the expression level. c Time course of relative ID1 gene expressions comparing arterial and venous endothelial cells under 1% oxygen conditions (n = 3 biological replicates). d Representative immunofluorescence images of arterial and venous endothelial cells stained for EPAS1 (purple) and nuclei (DAPI, blue) in normoxia and after 4 h of 1% oxygen exposure. Scale bar, 50 µm. e Quantification of the fold change of % EPAS1 colocalization with the nuclei over time comparing arterial and venous cells (n = 3 biological replicates, n = 3 field of view per replicate). f Venous endothelial tube formation assay under 21% and 1% oxygen conditions, both untreated and treated with PT2385. Quantification was based on the number of endothelial tube-like structures (n = 3 biological replicates). Scale bar, 400 µm. c, e, f Data points represent mean \(\pm\) s.d.; unpaired t-test comparing two independent groups (two-sided), one-way ANOVA for multiple comparisons (two-sided). Source data are provided as a Source Data file.