Protease Inhibition in Extracellular Trap Formation

To investigate whether proteases are required to form extracellular traps in coelomocytes, protease inhibitor cocktail (cOmplete Protease Inhibitor Tablets, Roche) was used as a broad protease inhibitor (BROAD PROT). It inhibits serine, cysteine, and metalloproteases in bacterial, mammalian, yeast, and plant cell extracts (CO-RO, Roche Mannheim, Germany) [54 (link)]. Additionally, we used [phenylmethlysulfonyl] fluoride (PMSF, Roche, Mannheim, Germany) that inhibits mainly serine proteases (1 mM, SERINE PROT) [55 (link)]. One milliliter of BROAD PROT contains: 375 μg/ml pancreas extract; pronase, 0.375 μg/ml; thermolysin, 0.2 μg/ml; chymotrypsin, 0.37 μg/ml; trypsin, 0.5 μg/ml; papain, 250 μg/ml (Roche, Mannheim, Germany). 10 μl of inhibitors were added to the cells in vitro or earthworms (the in vivo setting) were injected with 20 μl.