Fig. 2
Neutrophils pre-treated with excretory/secretory products exhibit reduced NET formation and altered nuclear morphology. The formation of NETs is reduced, accompanied by alterations in nuclear morphology. a, b The nuclear morphology of neutrophils (4 × 105) was untreated (a) or treated with 25 μM ATA alone (b). c, d 4 × 105 neutrophils were stimulated with 100 nM PMA for 3 h to induce NET formation or preincubated with 10 ng/μl ESP (added ATA) for 1 h before stimulation with PMA. Neutrophils were stained with Sytox green (green, extracellular DNA) and Hoechst 33342 (blue, total DNA) dyes (magnification 20×). The boxes show a close-up of the area indicated with white dotted lines. e Extracellular DNA quantification by fluorometry. ***P < 0.001. f Nuclear area quantification was carried out using ImageJ software. ATA, aurintricarboxylic acid; ESP, excretory/secretory products; PMA, phorbol-12-myristate-13-acetate. NETs, neutrophil extracellular traps