Phenotyping NK Cell Receptors

Analysis of activating receptors on NK cells were performed on PB and BM samples using a gating strategy on CD138^-CD45⁺CD56⁺CD3^-CD16^+/− cell after CD138⁺ cell (corresponding to PCs) gate exclusion (Figure 1A). The samples were stained with anti-CD138/FITC, anti-CD3/APC-H7, anti-CD56/PE, anti-CD45/PE-Cy7, anti-CD16/PerCP, anti-NKG2D/APC, anti-DNAM-1/FITC (BD Biosciences, San Jose, CA, USA) and anti-NKp30/APC (BioLegend, San Diego, CA, USA) for 25 min at 4 °C [50 (link)]. Expression levels of NK cell receptors on NK cell subsets derived from healthy PB donors are shown in Figure S4.
All the samples were acquired using a FACSCanto II (BD Biosciences, San Jose, CA) and data analysis was performed using the FlowJo 9.3.2 program (TreeStar, Ashland, OR, USA).