Fig. 6: Quantitative proteomic analysis of CD63-eGFP-RUSH and CD9-eGFP-RUSH EVs.

EVs were isolated by anti-GFP immuno-isolation from either non-transfected HeLa cells, or HeLa transfected with CD63-eGFP-RUSH or CD9-eGFP-RUSH, either 3 h or 24 h after biotin addition, and their composition was analyzed by mass-spectrometry. a Volcano plots representing quantified proteins with at least 2 peptides in 2 replicates in at least one condition. Shown are the fold changes of peptide abundancy between CD63- and CD9-eGFP expressing EV samples and the p-value of this quantification, for EVs recovered 3 h (left) or 24 h (right) after biotin addition. Position of the membrane-associated proteins selected for further analysis is indicated. b Results of the FunRich gene enrichment analysis among the proteins either enriched in the CD63- (blue), or CD9-eGFP (red) samples, or common between the CD63 and CD9-eGFP samples (purple) at 3 h or 24 h after biotin addition. For each subcellular compartment protein list, % of proteins of this category in the list of CD63-, CD9-, or common CD63/CD9 proteins is indicated, and the p-value of this percentage being different to its counterpart in the whole HeLa cell database is calculated. P-value (hypergeometric uncorrected). c Schematic representation of the transmembrane proteins identified in the CD63-, CD9-, or CD63/CD9-eGFP EVs at 3 h and 24 h.