Figure 1: Tamoxifen promotes extracellular trap formation in human neutrophils in vitro. | Nature Communications

Figure 1: Tamoxifen promotes extracellular trap formation in human neutrophils in vitro.

From: Tamoxifen augments the innate immune function of neutrophils through modulation of intracellular ceramide

Figure 1

(a) The effect of 10 μM tamoxifen (TAM; upper well) on chemotaxis in the presence or absence of 100 nM fMLP (lower well) was determined using a transwell system (n=12). (b) Extracellular DNA was quantified to determine the effect of both TAM and 4-hydroxytamoxifen (4-OHT) on NET release (n=9). (c) NET production was confirmed by fixing and immunostaining TAM-/4-OHT-stimulated cells with DAPI and a primary antibody for myeloperoxidase (blue: DAPI; green: NETs/myeloperoxidase); images representative of three independent experiments shown (scale bar=50 μm). (d) DCF-based ROS assays were performed to evaluate the effect of TAM and 4-HT on ROS production (n=9). (e) Neutrophils were pre-incubated with the ROS scavenger BHA before addition of either PMA to TAM to determine whether ROS production is required for tamoxifen-induced NET production (n=9). Where applicable, results were analysed by one-way analysis of variance and post hoc Newman Keuls test. *P<0.05, ***P<0.001 versus control values. BHA, butylated hydroxyanisole; DAPI, 4′,6-diamidino-2-phenylindole; NS, not significant; ROS, reactive oxygen species.

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