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. 2012 Dec 12;110(2):E141–E150. doi: 10.1073/pnas.1214361110

Fig. 3.

Fig. 3.

NICD induction rescues hematopoiesis from Hh inhibition in EBs. (A) Immunoflurescence showing Notch downstream target Hes1 (red) and DAPI (blue) in VE-cadherin+ cells sorted from day 4 EBs and treated with IHH, DAPT, or both for 36 h. Lower magnifies representative clusters that are highlighted with red arrows in Upper. (B) Box plot for the number of cells in each VE-cadherin+ cluster shown in A. (C) NICD expression in whole EBs on dox induction (n = 2). (D) CFUs from day 6 whole EBs that were NICD-overexpressed and/or cyc-treated during days 2–5 of differentiation (n = 4). (E) Flow cytometric quantification of VE-cadherin and CD41 populations from day 6 whole EBs that were NICD-overexpressed and/or cyc-treated during days 2–5 of differentiation (n = 2). (F) Blast and core colony formation during NICD overexpression and/or cyc treatment as assessed by BL-CFC assay (n = 3). (G) CFU potential of sorted VE-cadherin+ cells from day 6 EBs that were NICD-overexpressed and/or cyc-treated during days 2–5 of differentiation (n = 5). (H) Images of cells from hemato-endothelial culture at 10× objective (Upper) and flow cytometry for CD45+ cells from hemato-endothelial culture (Lower). Whole EBs were NICD-overexpressed from day 2 to 5; then, VE-cadherin+CD41−CD45− cells were sorted on day 6 and cultured in hemato-endothelial medium for 4 d.