Extended Data Figure 1: Induction of haemogenic endothelium from hPSCs.

a, Embryoid bodies formed from hPSCs. Subsequent panels show FACS analysis of day 8 embryoid bodies before magnetic cell isolation. b, CD34⁺FLK1⁺ population indicates haemogenic endothelium cells. CD235A and FLK1 plots show percentage of FLK1⁺CD235A⁻ cells in embryoid bodies, further gated with CD43 and CD34 plots to detect haemogenic endothelium (CD34⁺FLK1⁺CD235A⁻CD43⁻). c, d, Haemogenic endothelium isolated at day 8 (Supplementary Fig. 1) was further cultured in EHT medium for the indicated number of days. qRT–PCR showed (c) downregulation of endothelial genes and (d) upregulation of haematopoietic genes. HSPC genes (RUNX1, SCL/TAL1) peak on day 3 of EHT culture; consequently, this time point was chosen for introducing transcription factors followed by transplantation in subsequent experiments. e, Microscopy and FACS analysis on day 7 of EHT showed the appearance of haematopoietic cells (CD34⁺CD45⁺). Data shown as mean ± s.d.