Reply to Liu: The disassembly of the actin cytoskeleton is an early event during NETosis.

We used quantitative, high-resolution live-cell imaging of neutrophil-like human cells (dHL60) and mouse and human blood-derived polymorphonuclear neutrophils (PMNs) stimulated with ionomycin, lipopolysaccharide (LPS), or Candida albicans to identify 13 cellular events occurring during neutrophil extracellular trap formation (NETosis) (1). Following thousands of single cells over time showed that these events occurred in a specific order, with actin disassembly preceding all other cellular events observed, indicating that actin disassembly is an early event in NETosis (1).\n\nIn their letter to the editor, Liu challenges our conclusion (2). Liu uses flow cytometry of PMA-stimulated cells stained with “RFP-labeled phalloidin” to show that, at the population level, phalloidin fluorescence increases 60 min after addition of PMA and returns to basal level at 180 min, concluding that actin disassembly is not an early event of NETosis (2).\n\nThe points below support our published contention that … \n\n[↵][1]1To whom correspondence may be addressed. Email: watermancm{at}nhlbi.nih.gov.\n\n [1]: #xref-corresp-1-1