Fig. 3

Insulin withdrawal enhanced specification of arterial endothelium and inhibited hemogenic formation. a Schematic differentiation protocol and sample collection by flow cytometry. b Heatmap analysis of venous (blue), arterial (red), pan-endothelial (green), hematopoietic (yellow) and mesenchymal (gray) genes in day 5 and day 8 sorted CD34⁺CD31⁺CD43⁻ fractions in presence or absence of insulin, respectively. Two replicates performed in each group. c GO analysis of top differential upregulated genes in day 5 and day 8 EC fractions in absence of insulin. d Representative flow cytometry of day 7 CD184 and CD73 expression in CD31⁺CD34⁺ gated population in presence and absence of insulin, respectively. e Bar graph of percentage of CD184 and CD73 expression in CD31⁺CD34⁺ gated population in presence and absence of insulin on day 7, respectively. N > 3 independent biological replicates. f RT-qPCR analysis of CD34^hiCD43⁻ and CD34^med43⁺ subfractions from differentiating day 5 to day 8. HE hemogenic endothelium, HSPC hematopoietic stem/progenitor cell