Cd7 124 1d1

Cells were thawed at 37 °C, washed with sterile PBS and incubated with biotinylated anti-human lineage antibodies directed against CD2 (clone RPA-2.10, BioLegend), CD3 (HIT3a, 300304, BioLegend), CD4 (RPA-T4, BioLegend), CD7 (124-1D1, eBioscience), CD8a (RPA-T8, BioLegend), CD10 (SN5c, eBioscience), CD11b (ICRF44, BioLegend), CD14 (HCD14, BioLegend), CD19 (HIB19, BioLegend), CD20 (2H7, eBioscience), CD56 (HCD56, BioLegend) and GPA (HIR2, BioLegend) and LIVE/DEAD Fixable Aqua Dead Cell Stain (L-34957, Life Technologies). For gating on hematopoietic progenitor cells, this was followed by secondary staining with anti-human CD34–APC (8G12, BD Biosciences), anti-CD38–PE/Cy7 (HIT2, BioLegend), anti-CD90–FITC (5E10, BD Biosciences), anti-CD45RA-PB (MEM-56, Thermo Fisher Scientific), anti-CD123–PE (7G3, BD Biosciences) and streptavidin-APC/Cy7 (BioLegend). Intracellular staining with anti-human Ki-67–BV605 antibody (BioLegend) was performed using the BD Cytofix/Cytoperm Fixation/Permeabilization kit. Samples were run on an LSR II flow cytometer (BD Biosciences), and recorded events were analyzed with FlowJo 10 software (BD). Individual fluorescence-minus-one controls were used to determine gating. HSCs were identified as lineage⁻CD34⁺CD38⁻CD45RA^loCD90⁺ cells, CMPs as lineage⁻CD34⁺CD38^intCD45 RA⁻CD123^int cells and GMPs as lineage⁻CD34⁺CD38^intCD45RA⁺CD123^int cells.