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

Figure 7

Role of intracellular calcium in NETosis induced by a cocaine-levamisole mixture. Neutrophil-enriched suspensions from healthy controls were incubated with BAPTA at different concentrations for 10 min before adding levamisole (40 nM) and the cocaine-levamisole mixture (20 µM/40 nM) for 3 h. After incubation, cells were stained with Sytox Green and Hoechst 33342, washed, and analyzed by flow cytometry. (a) Representative dot plots showing double positive Sytox Green⁺ Hoechst 33342⁺ netting neutrophils in presence of BAPTA. (b) Representative dot plots showing the effect of 2 µM BAPTA on reduction of double positive Sytox Green⁺ Hoechst 33342⁺ netting neutrophils induced in the presence of the cocaine-levamisole mixture or levamisole. n = 2 independent experiments.

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(b)