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. 2018 Jun 29;9:1514. doi: 10.3389/fimmu.2018.01514

Figure 1.

Figure 1

Prostratin (PRO) and bryostatin-1 (BRY) activate natural killer (NK) cells and modulate activating receptors. (A) The purity of NK cells isolated from PBMCs was examined by flow cytometry measuring the frequency of CD3−CD56+CD16+/− cells. (B) NK cells were cultured for 18, 46, and 76 h in medium alone [not treated (nt)] or supplemented with 1 or 10 µM PRO, or with 5 or 10 nM BRY. The viability of gated NK cells was examined by LIVE/DEAD staining and expressed relatively to nt cells set at 100%. Bars represent mean ± SEM (n = 3). (C–F) NK cells treated or not with 1 µM PRO, 10 nM BRY, or 12.5 ng/ml of IL-15 were examined at various time points as indicated (C,D,F) or after 18 h only (E) for the expression of various markers: (C,D) the frequency of CD69+ and CD107a+ NK cells among nt (filled gray histograms, gray percentages) and treated cells (open histograms, black percentages) is shown together with control IgG signal (dashed line) for a representative experiment (C) and as mean ± SEM (D); (E,F) mean ± SEM of both percentage of positive cells and mean fluorescence intensity (MFI) for NKG2D, DNAM-1, NKp46, NKp44, NKp30, and CD16 is shown. Experiments were performed with at least three independent donors [up to eight in panel (E)]. *P < 0.05, **P < 0.01, and ***P < 0.001 by paired t test.