Fig. 3.

Representative gating strategy for NK-cells. A) Lymphocytes were identified in a SSC-A × FSC-A plot prior to the removal of doublets in a FSC-H × FSC-A plot. Next, a fixable viability stain (FVS) was used to remove non-viable cells prior the removal of monocytes using a histogram. B) In a pre-exercise sample, CD56 was plotted against CD3 to identify total CD3⁻CD56⁺ NK-cells. In the CD3⁻CD56⁺ NK-cell population: CD56^dim and CD56^bright NK-cells were identified in a FSC-A × CD56 plot; NK-cells capable of, and susceptible daratumumab-mediated ADCC were identified in a CD38 × CD16 plot; and CD57⁻ and CD57⁺ NK-cells were identified using a histogram. CD3⁻CD56⁺CD57⁻ and CD3⁻CD56⁺CD57⁺ NK-cells were further gated to identify CD16 sub-populations. C) Represents the same gating strategy as ‘B)’ but from a post-exercise sample. SSC-A, side scatter-area; FSC-A, forward scatter-area; FSC-H, forward scatter-height; SMM, smouldering multiple myeloma.