Extended Data Fig. 5. E-selectin stimulation does not result in granule release.
Enrichment of granule content (azurophilic gr., specific gr., gelatinase gr.) in supernatants of E-selectin (1 µg ml-1) and PBS (control) stimulated human neutrophils was analyzed by mass spectrometry (n = 3 independent experiments). a Volcano plot showing differentially released proteins, cytosolic small alarmins (Mw≤50 kDa), cytosolic alarmins (Mw>50 kDa) and granules are indicated. b Release of indicated proteins for each donor, c release of all proteins, all granules, azurophilic granules, specific granules and gelatinase granules. d-g Isolated human neutrophils were stimulated with E-selectin (1 µg ml-1), PMA (100 nM, positive control) or PBS (control) for 10 min. Surface expression of d CD63, e CD66b and f CD11b was assessed using flow cytometry (n = 4 independent experiments). g gating strategy. Data are presented as mean ± s.e.m. (one-way ANOVA, Tukey’s comparison for d-f, as volcano plot for a, as heat map for b, as box-whisker-plots for c (50% IQR, median center, whiskers ranging from 5-95 percentiles and dots indicating outliers), as representative flow cytometry dot blot image for g, and as representative histograms for d-f. -log10 transformed p-values depicted in the volcano plots were determined by Welch’s two-sided t-test. Enrichment of granule content in c was determined by a 1D annotation enrichment with Benjamin-Hochberg FDR = 0.02. ns: not significant.