Insight into Neutrophil Extracellular Traps through Systematic Evaluation of Citrullination and Peptidylarginine Deiminases

Figure 2

Induction of NETs in murine neutrophils. Murine neutrophils were left untreated (UN) or were treated with ionomycin (IO), MSU, PMA, and C. albicans (CA), fixed, and stained with DAPI (blue) and anti-citrulline antibody (pink). Image labeled “Secondary” was created by stimulating neutrophils with ionomycin and staining without the F95 primary antibody and only the anti-mouse IgM-TRITC secondary antibody as a negative control. (a) Representative images at 400x, . Enlarged insets demonstrate donut-like structures (doNETs). The number of neutrophils and NETs were quantified. Graphs depict the average and SEM for percent of neutrophils that formed total NETs (b), citrullinated NETs (c), and uncitrullinated NETs (d) for each condition with percent NETs for each stimulant compared to untreated. (e) The percent of citrullinated versus uncitrullinated NETs was compared for each stimulus with average and SEM graphed. For all panels: ; , , and .

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