Figure 2.

Acquisition of Endothelial and Hematopoietic Markers Occurs Normally upon hMLL1 Induction

(A) Flk-1 surface expression during differentiation of EBs. One representative of three independent experiments is shown as average live-gated, Flk-1⁺ cells ± SEM of quadruplicate cultures. hMLL1-inducible EBs were treated with vehicle (PBS) or 2 μg/mL doxycycline for indicated days.

(B) Generation of hemangioblast-enriched cells shown by flow cytometry. hMLL1i EBs were treated with solvent (PBS) or doxycycline from day 2 to 4. Quadrant gating is based on single stained controls and the hemangioblast-enriched Flk-1⁺/Pdgfrα^neg population is indicated in red and quantified in the bar graphs (right). Data are representative of two independent experiments and are shown as the average ± SEM of triplicate cultures.

(C and D) (C) Hemogenic endothelium c-Kit⁺/Tie-2⁺ and (D) hematopoietic Cd41⁺/Cd45⁺ were determined and quantified in the bar graphs. hMLL1i EBs were treated with solvent (PBS) or doxycycline from day 4 to 7. Data are representative of three independent experiments and are shown as averages ± SEM of triplicate cultures.