Whole BM flush were collected from tibia and femur into buffer containing PBS [3% (v/v) bovine serum albumin (BSA)] and filtered through 40-μm nylon filter (BD Biosciences, San Jose, CA). Red blood cells (RBCs) were lysed with RBC lysis buffer (Thermo Fisher Scientific) for 5 min at RT. Cells were stained with CD39 phycoerythrin (PE)/cyanine-7 (8 μg/ml; 143805, BioLegend, San Diego, CA), CD73 PE (1.25 μg/ml; 12-0731-82, Thermo Fisher Scientific), and CD45 allophycocyanin (APC) (1.25 μg/ml; 17-0451-82, Thermo Fisher Scientific) antibodies for 30 min at RT. Stained cells were analyzed with BD Accuri C6 flow cytometer and CFlow software. Analyses were performed by comparing to unstained cells and gated for singlets.

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