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Cells were washed twice with the FLICA kit wash buffer and then incubated with LIVE/DEAD Fixable AQUA Dead Cells Stain (Thermo Fisher, USA) for 15 min at RT, followed by extracellular staining in PBS + 1% BSA with the following fluorochrome-conjugated antibodies for monocyte phenotyping: anti-CD14 BV605 [Clone: M5E2], anti-CD16 PE-Cy7/BV711 [Clone: 3G8], anti-CD3 PE [Clone: HIT3a] and anti-CCR2 PerCP Cy5.5 [Clone: K036C2] are from BioLegend

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The product of this process is primarily the hydroxy free radical, the most poisonous of all ROS

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