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In some experiments, cell death was analyzed by flow cytometry using human neutrophils and the probe Zombie violet (BioLegend, 423113), an amine-reactive fluorescent dye that is nonpermeant to live cells but permeant to the cells with compromised membranes

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The beads were washed twice with 500 l of equilibration buffer and twice with 500 l of storage buffer (18 mM HEPES pH 7.4, 100 mM NaCl, 1 mM MgCl 2 , 5% (v/v) glycerol and 0.003% (v/v) IGEPAL CA-630), before incubation in 100 l of SENP EuB elution solution (500 nM of in-house purified SENP EuB protease 51 in storage buffer) at 4 C with gentle rotation for 1 h