The protein Tsg101 plays an integral role extracellular vesicle formation and secretion from cells. We developed a mouse model with Tsg101 knockdown (Tsg101dAd). Visceral adipose tissue (VAT) was harvested from Tsg101dAD and Tsg101flfl mice fed high-fat diet to induce obesity or matched diet as non-obese controls. VAT was ex vivo cultured for 48 hrs and its conditioned media was collected. Extracellular vesicles were isolated from conditioned media by ultracentrifugation. The aim of the project was to characterize extracellular vesicle protein cargo from obese and non-obese VAT.