Neutrophils of healthy donors were stimulated using PMA (25 nM) for 2.5 h, and the impermeable DNA dye SYTOX Green (Thermo Fisher, USA, Cat No. S7020) was added to each well at a volume ratio of 1 : 100. Subsequently, fluorescence intensity was measured using a SpectraMax M5 multifunctional microplate reader (Biotek, USA) 10 min later. After PMA stimulation, the fluorescence intensity was recorded every 30 min for a total of 3.5 h.
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