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For chemical induction of NETs, neutrophils were treated with PMA (100 nM) for 6 h, LPS (10 μg ml −1 ) for 2 h or A23187 (1.25 μM), nigericin (5 μM), zymosan (25 μg ml −1 ) and MSU (100 μg ml −1 ) for 4 h in a cell culture incubator