NET proteome assembly is different in PVL-induced NETs. (A) Principal component analysis of the proteomes of naïve neutrophils, and PVL-induced (10 nM), PMA-induced (50 nM), and nigericin-induced (15 µM) NETs. Analysis was performed on scaled log2-abundance of all proteins detected in at least 2 of 3 replicates. (B) Euler diagram showing the number and distribution of significant differentially abundant peptides (logFC > 1, adjusted P < 0.01) on NETs compared with naïve neutrophils. Areas are proportional to DAP set size. (C) Clustered heatmap showing fold change (log2) values on each NET sample compared with naïve neutrophils of significant DAPs from panel B. Clustering was performed by k-means algorithm with k = 4 clusters. (D, E) Top 25 most abundant DAPs that are significantly differentially abundant on PVL-induced NETs compared with PMA-induced (D) and nigericin-induced NETs (E). Point size and fill color represent average abundance across samples and adjusted P value, respectively. Proteomes were made from n = 3 samples per condition from independent donors. DAP significance for each comparison was determined by a threshold of |[log2 fold change]| > 1 and adjusted P value < 0.01.