Endothelial cell dysfunction plays a critical role in the pathogenesis of chronic allograft dysfunction and tissue fibrosis. Autophagy deficiency in endothelial cells has been implicated in altered extracellular vesicle (EV) cargo composition, which may contribute to pericyte-to-myofibroblast transition. In this study, we performed quantitative proteomic analysis of EVs isolated from the culture supernatant of mouse aortic endothelial cells (MAECs) treated with or without TGF-β to identify differentially expressed proteins. EVs were isolated by ultracentrifugation and characterized according to MISEV2023 guidelines. LC-MS/MS-based proteomics was performed to compare the protein profiles between control (NC) and TGF-β-treated groups. This dataset provides insights into the molecular cargo alterations in endothelial cell-derived EVs under pro-fibrotic conditions.