Figure 12.

NETosis induced by E. histolytica trophozoites is independent of PAD4 activity. (A) Neutrophils (3 × 10⁵) were pre-incubated with 10 μM GSK484 or DMSO for 30 min and then stimulated with 20 nM PMA, 10 μM A23187, 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, fixed and stained with DAPI. Images were taken at 60x magnification. Scale bar 50 μm.