osomes and lysates listed were probed with anti-mouse and anti-rabbit secondary antibodies only. Molecular weight markers are indicated at the sides of the blots. Exosome surface HSP90 was identified by fluorescence activated cell sorting analysis of exosomes bound to latex beads and treated as if PubMed ID:http://www.ncbi.nlm.nih.gov/pubmed/22203832 they were cells in FACS. Gray fill indicates fluorescence of exosome-coated beads probed with a fluorescently-labeled isotype control antibody, and the red line shows fluorescence intensity of the exosome/bead complex with the fluorescentlylabeled anti-HSP90 antibody. doi:10.1371/journal.pone.0042064.g002 identified and actual proteins being studied. As a caveat, it must be stated that numerous proteins we identified have multiple subcellular localizations, particularly in tumors; for example, the chaperones of the HSP70 and HSP90 families, as well as HSP27 and protein disulfide isomerase, may translocate to the nucleus, the cytoplasm, and even the cell surface. The proteins run the gamut of activities and functions, including cytoskeletal and structural components, nucleic acid-binding proteins, transcriptional and translational regulators, transporters, chaperones, kinases and signaling components, and a wide variety of enzymes. Functionally, the largest single category of proteins could be grouped as enzymes, with nearly the same percentage as seen previously. Transcriptional regulators, transport proteins, and structural proteins combine for over one-third of the remaining functions, with chaperones, nucleic acid binding proteins, scaffold proteins and proteins of unknown function holding similar percentages. The lowest represented functions were proteases/inhibitors, translational regulators, motor proteins, kinases, and hormones. A similar caveat applies in that many of these proteins are multifunctional and may play multiple roles, particularly in complexes, thus making definitive categorization difficult. Using NU-7441 chemical information Integrated Pathway Analysis software, the identified proteins were grouped into networks of associated functions, canonical pathways, and disease and toxicology relationships. The top 5 networks/associated functions were ��Cell Morphology, Post-Translational Modification, Protein Folding”; ��Genetic Disorder, Hematological Disease, Renal and Urological Disease”; ��Carbohydrate Metabolism, Energy Production, Nucleic Acid Metabolism”; ��Neurological Disease, Genetic Disorder, Hematological Disease”; ��Carbohydrate Metabolism, Gastrointestinal Disease, Genetic Disorder”. The scores reflect the probabilities of such associations occurring by chance, with the threshold value for significance set at 1.25; as evident, the scores are highly significant. Medulloblastoma Exosome Proteomics Suggest Exosome Functions Functional Roles of Medulloblastoma Exosomes 6 Functional Roles of Medulloblastoma Exosomes promote tumor cell migration in a concentration-dependent fashion which is minimally as good as, or better than, FBS and at higher 
exosome concentrations is significantly better. Whether the cells were serum-starved or not is of no consequence in terms of baseline migration. The migration of D283MED cells through what was essentially naked plastic towards exosomes is impressive given the lack of adherence of these cells to commonplace matrix substrates. Adherent medulloblastoma cell lines UW228 and DAOY also migrate towards cognate exosomes in a dose-dependent fashion. The merged interactomes of Networks 3 and 8 focus on a number of immu
