Human PBMCs were stained with anti-CD14 (clone M5E2, Biolegend), anti-CD16 (clone 3G8, Biolegend), anti-CD4 (clone OKT4, Biolegend), human CD39 (clone 498403, R&D Systems), human CD73 (clone 606112, R&D Systems)44 (link),45 (link), human CD25 (clone BC96, Biolegend), human FoxP3 (clone 236 A/E7, Biolegend), and human PD-1 (clone EH12.2H7, Biolegend). Prior to anti-FoxP3 staining, the cells were fixed and permeabilized.
Stained cells were analyzed in the Oklahoma Medical Research Facility (OMRF) Flow Cytometry Core Facility on a BD LSRII (BD Biosciences) and data was analyzed using FlowJo Software (Tree Star, Inc., Ashland, OR). Single stained samples were used to determine compensation values, and fluorescence minus one (FMO) controls were used to determine gating placement.
Stained cells were analyzed in the Oklahoma Medical Research Facility (OMRF) Flow Cytometry Core Facility on a BD LSRII (BD Biosciences) and data was analyzed using FlowJo Software (Tree Star, Inc., Ashland, OR). Single stained samples were used to determine compensation values, and fluorescence minus one (FMO) controls were used to determine gating placement.
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