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. Author manuscript; available in PMC: 2015 Nov 20.
Published in final edited form as: Chem Biol. 2014 Nov 20;21(11):1457–1462. doi: 10.1016/j.chembiol.2014.09.015

Figure 2.

Figure 2

N-AgrD peptide variants stimulate PMN chemotaxis and NET formation. (a) As an indicator of migration, elastase enzymatic activity in neutrophil supernatants was measured at 405 nm. Formylated peptides N-AgrD F20 and N-AgrD F24 stimulated increased chemotaxis. The N-AgrD D20 peptide, which is non-formylated, did not exhibit chemoattractant properties. (b) PMN calcium influx indicating receptor activation. A significant decrease in activation was observed by each pharmacological blockade, indicating the N-AgrD peptide family functions though a formyl peptide receptor-dependent mechanism. BI = BOC, selective inhibitor of FPR1; PI: PBP, selective inhibitor of FPR2. (c) The Quant-iT Picogreen assay was used to quantify extracellular DNA release upon stimulation by the N-AgrD peptides. Only formylated peptide NAgrD F20 stimulated a significant increase in NET production. (d) Microscopy confirmed only peptide N-AgrD D20 stimulated formation of web-like DNA structures similar to the PMA positive control. Statistical analysis by one-way ANOVA; *** P <0.001. Data was expressed as mean ± standard deviation.