ICN1 induction promotes VE-cadherin⁺ hemogenic endothelial cell specification. (A) Flow cytometric analysis of day 6 iICN1 EBs with (ICN 3-5) or without (ICN 0) Dox induction (0.5 μg/mL) from days 3 to 5 is shown. (B) CD45⁻ VE SP cells were sorted from day 6 iICN1 EBs with (ICN 3-5) or without (ICN 0) Dox induction (0.5 μg/mL) from days 3 to 5, and the same number of sorted cells was subjected to HE culture. Flow cytometric analysis of day 4 HE culture is shown. (C) The cumulative average of CD45⁺ cells of days 2, 4, and 6 HE culture is shown. n = 3; *P < .05 by Student t test in comparison with noninduced control. (D) Cytospins of day 10 HE culture of CD45⁻ CD41 SP, VE SP, and DP cells from day 6 iICN1 EBs with (ICN 3-5) or without (ICN 0) Dox induction (0.5 μg/mL) from days 3 to 5 are shown. ICN1 was not induced in HE culture. Representatives of 3 independent experiments are shown.