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Collected cells were centrifuged at 1000 rpm for 5 mins and resuspended in 1mL cold FACS Buffer (3 % PFA + 0.2 % BSA, in 1x PBS)

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Also, blocking autophagy with 3-MA or bafilomycin A1 increased NETs formation in PMA-stimulated neutrophils, while enhancing autophagy with trehalose decreased NETs formation in LPS-stimulated neutrophils