All antibodies used for flow cytometry are detailed in Supplementary Table 1 ).
To prepare cells for FACS, all cells were recovered for 20 minutes at 37°C in the presence of 200 U/ml DNase (Worthington Biochemical, Lakewood, NJ) in IMDM with 10% fetal bovine serum. After recovery, viable mononuclear cells were separated by a Ficoll density gradient (GE Healthcare). When necessary, CD34-based enrichment was performed using paramagnetic MACS beads (Miltenyi Biotech Inc, San Diego, CA) per the manufacturer’s protocol.
FACS sorting was performed on a Becton Dickinson FACSAria II. All cells were resuspended in and sorted into cold FACS Buffer (PBS + 2% FBS + 2 mM EDTA) containing propidium iodide at 1 ug/ml or 4′,6-diamidino-2-phenylindole (DAPI) at 1 ug/ml. All cell sorting steps were validated using post-sort analyses to verify purity of sorted cell populations (Supplementary Table 1 ).
To prepare cells for FACS, all cells were recovered for 20 minutes at 37°C in the presence of 200 U/ml DNase (Worthington Biochemical, Lakewood, NJ) in IMDM with 10% fetal bovine serum. After recovery, viable mononuclear cells were separated by a Ficoll density gradient (GE Healthcare). When necessary, CD34-based enrichment was performed using paramagnetic MACS beads (Miltenyi Biotech Inc, San Diego, CA) per the manufacturer’s protocol.
FACS sorting was performed on a Becton Dickinson FACSAria II. All cells were resuspended in and sorted into cold FACS Buffer (PBS + 2% FBS + 2 mM EDTA) containing propidium iodide at 1 ug/ml or 4′,6-diamidino-2-phenylindole (DAPI) at 1 ug/ml. All cell sorting steps were validated using post-sort analyses to verify purity of sorted cell populations (