Fig. 1

Frequency of responding NK cells across multiple stimulation conditions. a Top row: overall successive gating strategy demonstrates initial broad gating on forward and side scatter to include large lymphocytes. Forward area and height are used to discriminate single cells followed by identification of viable cells. Monocytes and B cells are excluded using CD14 and CD19, and T cells are excluded using CD3 and CD4. Bottom row: CD56 and CD16 are used to identify NK cells and changes in CD16 expression across stimulation conditions. Black gate is the total NK population and red gate is CD56 dim population. b A univariate analysis of the frequency of CD107a, INF-γ, TNF, granzyme B, perforin, and eomesodermin expression from the total NK cell population as well as CD57 and CD8 expression are shown. c Pie charts of classic Boolean analysis of five functional markers measured (CD107a, IFN-γ, TNF, granzyme B, and perforin). Comparison of pies and nonparametric partial permutation tests (Monte Carlo simulation) are performed to determine statistical significance between pies (p-value < 0.0001)⁶⁹. d Graph of classic Boolean analysis of five functional markers measured. Four healthy subjects from four independent experiments in each stimulation condition are included. CD107a, IFN-γ, and TNF are mock subtracted (i.e., stimulated condition − unstimulated result). Black bars represent the standard error mean (SEM) for each stimulation condition calculated as the variance of the sampling distribution obtained, equal to the variance of the population divided by the sample size