Fig. 2

Proteomic Comparison of Ultracentrifugation versus CD9 Affinity purification for Isolation of Plasma or Ascites Extracellular Vesicles. A Nanoparticle tracking analysis of ascites EVs purified by UC or CD9AP demonstrates a subset of EVs are enriched by CD9AP. The distribution of CD9AP-EVs were primarily distributed around < 150 nm in diameter, whereas UC samples were comprised of a heterogenous mixture of EVs that were primarily distributed around ~ 200 nm, albeit subpopulations of EVs were detectable up to 900 nm (see Table S3). B Heatmap of identified proteins and dendrogram demonstrate increased proteomic depth obtained by UC compared to CD9AP using paired ascites donors. C Cellular debris and large EV components, such as actin, were depleted using CD9AP. D 145 and 1953 proteins were exclusive to EVs enriched by CD9AP or UC (> 1 replicate), whereas 416 proteins were common to both CD9AP and UC-enriched EVs (> 2 replicates in each condition). E Volcano plot of common proteins to CD9AP and UC identified 64 and 84 proteins significantly enriched in either CD9AP- or UC-enriched ascites EVs, respectively. F 185 proteins were significant enriched within ascites EVs compared to blood plasma EVs collected by UC from healthy donors. G 55 proteins were significantly enriched using CD9AP on ascites EVs compared to blood plasma EVs collected from healthy donors