Anthracyclines suppress NETosis induced by both Nox-dependent and -independent pathway agonists. NETosis assays were performed on human neutrophils resuspended with 20 μM of each of the 126 anti-neoplastic compounds with 5 μM Sytox Green fluorescence dye. After one hour, the neutrophils were either unstimulated (-ve control) or stimulated using one of the four agonists PMA (Phorbol 12-myristate 13-acetate) or LPS (Lipopolysaccharides), A23187 or ionomycin). Fluorescence intensities (proxy for DNA release) were measured by a plate reader every 30 min for 4 h. At the last time point (4 h), % DNA release in each conditions were divided by the fluorescent intensity of the cells lysed by Triton-X (Triton lyses the cells and gives maximum fluorescence). Furthermore, the % DNA release (NETosis) in each compound was calculated by considering the 100% DNA release at the last time point (240 min) in respective agonists, as represented in the heatmap. Compounds are grouped by class/mechanism of action. The anthracyclines consistently show less DNA release in each condition.