Figure 7.

NETosis induced by trophozoites is independent of NOX2-derived ROS. (A) Neutrophils (3 × 10⁵) were pre-incubated with 200 μM apocynin or DMSO for 30 min and then stimulated with 20 nM PMA or co-cultured with trophozoites (1.5 × 10⁴) during 4 h. Supernatants were collected, mixed 1:1 with SYTOX Green (500 nM) and fluorescence was measured. NETs amount is expressed in fluorescence relative units (FRU). Values are means ± SD of three independent experiments in triplicate. ^*p < 0.001; NS: statistically non-significant. (B) Neutrophils were treated as describe previously, cells were fixed and stained with DAPI. Images were taken at 60x magnification. Scale bar 50 μm.