Fig. 1

Effect of ATA on neutrophils cell activity. a The cell viability of neutrophils was assessed under different ATA concentration treatments (5, 10, 15, 20, 25, and 30 μM) using the CCK-8 assay (n = 5). Different concentrations of ESP impede PMA-stimulated NET formation. b Unstimulated neutrophils were used as a control group. c Neutrophils were incubated with 25 μM ATA alone. d Neutrophils were treated with 100 nM PMA and 25 μM ATA. e–g Neutrophils were pretreated with Ad3 ESP at different concentrations (2.5, 5, and 10 ng/μl) containing 25 μM ATA for 1 h at 37 °C before incubation with PMA. NETs were visualized by staining with Hoechst 33342 (blue) and Sytox Green (green). Merged representative images of at least three experiments are shown. Scale bars = 80 μm. h Extracellular DNA quantification by fluorometry. ***P < 0.001. ATA, aurintricarboxylic acid; CCK-8, Cell-Counting Kit 8; ESP, excretory/secretory products; PMA, phorbol-12-myristate-13-acetate. NETs, neutrophil extracellular traps