Figure 1.

CQ and HCQ inhibit NETs induced with various NET activators. (a) Murine neutrophils were isolated from bone marrow using density gradient centrifugation, pre-treated with vehicle, CQ (1 mM), or HCQ (1 mM) then stimulated to form NETs with either PAF (50 µM) or LPS (10 µg/ml) for 2 h. DNA was stained with Hoechst (red) to visualize extracellular DNA and images were obtained to visualize NET structures at 20× using a Zeiss microscope. Both CQ and HCQ led to a subjective decrease in visualized extracellular DNA, consistent with reduced NET formation. (b and c) NET formation was quantified by measuring cf-DNA released into the supernatant with QuantiT Pico Green kit per the manufacturer’s protocol. CQ and HCQ led to a significant reduction in cf-DNA in both PAF and LPS-induced NET assays.*P value < 0.05.