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To determine the K M of NADH in the forward reaction, NADP + was held constant using glutathione reductase and oxidized glutathione (0.5 mM NADP + , 4 U/ml glutathione reductase, 1 mM oxidized glutathione and various concentrations of NADH) whereas determination of K M for NADP + was obtained using alcohol dehydrogenase to hold NADH concentrations constant (50 M NADH, 1.2 U/ml alcohol dehydrogenase, 130 mM ethanol, 6 mM hydrazine (pH 7.0) and various concentrations of NADP + )

These findings confirm the functional loss of XKR8 in neutrophils from XKR8 KO mice and support its essential role in both apoptotic and NETosis-associated lipid scrambling