Fig. 2

Nasal exosomes induce immune cell migration and the use of exclusion lists during mass spectrometry identifies new proteins and peptides. a Human monocytes, NK cells and neutrophils isolated from blood were added to one of the chambers of a Boyden chamber (35,000–250,000 cells/well). To the other chamber 30 µl of the different doses of nasal exosomes were added. Media was used as a control. After five (neutrophils) or 12 (monocytes and NK cells) hours the number of cells migrated to the exosome-containing chamber on the other side of the membrane were analysed. Kruskal–Wallis test followed by Dunn’s multiple comparisons test were used to determine significant differences where all concentrations were only compared to the control. P values * <0.05, ** <0.01, *** <0.001, **** <0.0001. b The Venn diagrams compare the proteins identified in the first acquisition (black font), the second acquisition (red font) and the third acquisition (blue font) and shows that the utilisation of exclusion lists in re-acquisitions led to increased numbers of identified proteins. c The utilisation of exclusion lists also resulted in the identification of new unique peptides for proteins previously identified, which increased the coverage and confidence for these proteins. Proteins were divided into groups based on the number of peptides identified in the first acquisition for each protein (1–5 or more). Data are presented as the percentage of proteins identified with additional unique peptides in the second or third acquisition in each category