Canonical pathways altered by the EVs signature
The coordinated function displayed by the proteins contained in the anaphylactic panel would support information about the molecular bases of the reaction, especially of those exclusively found in the acute condition. Therefore, it was performed two comprehensive analyses: a “manual” based in the Uniprot database and an in silicoapproach through IPA. The first revealed a major group of immune proteins and those participating of the cellular structure, supporting a role of EVs in anaphylaxis. A summary of the major cell location as well as the main biological processes of identified EV markers is illustrated in Figures 4A-B . In agreement, the in silico IPA revealed different canonical pathways, candidates to underlie signaling in anaphylaxis. A list including those with higher significance is showed in Figure 4C . This analysis showed that these proteins altered involve mainly immunological processes such as degranulation of cells (mainly neutrophils) and adhesion-binding of leukocytes (Sup Table 2 ). A major interest was aroused by the appearance/repetition of same molecules participating in the different top canonical pathways; like CDC42. Therefore, to better visualize associations between the key proteins and their multiple participation in the functional networks described previously, a graphical representation based on STRING analysis was performed. As expected, this demonstrated that some AnEVs proteins are grouped in a unique cluster, such is the case of the keratinocytes include in the glucocorticoid receptor signaling pathway. In addition, Clathrin-mediated endocytosis, a process related with the own biology of EVs, was also observed.