NETosis Secondary to the Use of Levamisole-Adulterated Cocaine: A Likely Underlying Mechanism of Vasculopathy

Figure 3

Evaluation of MPO in NETs extruded from cultures of neutrophils exposed to cocaine, levamisole, and cocaine-levamisole mixture. Neutrophil-enriched suspensions from healthy controls were exposed to cocaine (20 µM), levamisole (40 nM), cocaine-levamisole mixture (20 µM/40 nM), and PMA (20 nM) for 6 h. Staining with Sytox Green, Hoechst 33342, and anti-MPO-PE and anti-NE-Alexa fluor 647 mAbs was used to evaluate extrusion of neutrophil DNA and granular content. (a) Representative dot plots and (b) column chart showing the percentages of double positive Hoechst 33342⁺ MPO⁺ netting neutrophils generated in response to each treatment. Bar represents median values and error bars correspond to the IQRs. ; Kruskal–Wallis test. n = 4 independent experiments.

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