Whole blood (100 µL) was incubated with 5 µL of V500 CD45 (BD Bioscience, San Jose, CA, USA), 20 µL of PerCP-Cy5.5 CD34 (BD Bioscience, San Jose, CA, USA), 5 µL of PE VEGFR-2 (R & D Systems, Minneapolis, MN, USA ), 5 µL APC CD133 (Miltenyi Biotic Inc., Bergisch Gladbach, Germany) and 10 µL of FITC CD144 (BD Bioscience, San Jose, CA, USA) for 30 min. Subsequently, 2 mL of PharmLyse (BD Bioscience, San Jose, CA, USA) were used to lyse the red cells. The sample was then analyzed by flow cytometry on a BD FACS Canto™ II system and the results data processed using BD FACSDiva™ software as described previously [14 (link)].
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