Cd303 percp cy5

Dendritic cell-enriched cell suspensions were stained with an antibody cocktail consisting of the following antibodies in PBS + 2% human sera for 30 min on ice: CD1c-APC/Cy7 (L161; BioLegend), CD3-BUV395 (UCHT1; BD Bioscience), CD11b-A700 (M1/70; BioLegend), CD11c-PE/Cy7 (3.9; BioLegend), CD14-A700 (HCD14; BioLegend), CD19-V450 (HIB19; BD Bioscience), CD20-eFluor 450 (2H7; eBioscience), CD56-BV421 (5.1H11; BioLegend), CD123–BV605 (6H6; BioLegend), CD141-BV711 (1A4; BD Bioscience), CD303-PerCP/Cy5.5 (201A; BioLegend), HLA-DR-PE-CF594 (G46-6; BD Bioscience), and NKp46-BV421 (9E2; BioLegend). After washing the cells, they were resuspended in PBS + 2% human sera + 0.1 mg/ml 4′,6-diamidino-2-phenylindole and cell sorted using a BD FACSAria II cell sorter into CD1c⁺ DCs (CD3⁻CD11b⁻CD14⁻CD19⁻CD20⁻CD56⁻CD123⁻NKp46⁻HLA–DR⁺CD11c⁺CD1c⁺), CD141⁺ DCs (CD1c⁻CD3⁻CD11b⁻CD14⁻CD19⁻CD20⁻CD56⁻CD123⁻NKp46⁻HLA-DR⁺CD11c⁺CD141⁺), pDCs (CD1c⁻CD3⁻CD11b⁻CD14⁻CD19⁻CD20⁻CD56⁻NKp46⁻HLA-DR⁺CD123⁺CD303⁺), and monocytes (CD3⁻CD19⁻CD20⁻CD56⁻CD123⁻NKp46⁻HLA-DR⁺CD11b⁺CD14⁺). The purity of sorted cell populations was reanalyzed and routinely above 95%.