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2 Harmonized NK FACS panels to study NK cell phenotype and function | 65 Supplementary figure 3: Gating strategy for the NK cell function panel The gating procedure to analyse NK cell functions from PBMC population is shown. Singlets were selected gating PBMC against FCS-A versus FSC-H. CD45+ cells negative for CD14, CD19 and SYTOX® Blue were further gated against CD3/TCRγδ and CD56 to differentiate NK cells, T cells and CD3+CD56+ cells. CD45+CD56+ NK cells were further analysed for CD16, CD107a, NKp44 and CD25 expression. NK cells shown in this figure were stimulated with A431 + CET to show higher CD107a and lower CD16 levels. Further to facilitate NK subset analysis, NK cells were sub-divided into NK bright and NK dim populations based on CD56 expression and CD16 levels. CD45+CD56- cells can also be studied using this gating strategy.