We have employed the stack gel method to analyze the proteomic content of EVs. Briefly, 50 ug of EVs sample was lysed using RIPA buffer with 1 mM PMSF for 10 minutes in ice with occasional vortexing. peptide solution was transferred to the tube containing 50 mM ammonium bicarbonate and the step with 50% v/v acetonitrile, 5% formic acid was repeated again. Following this, the peptide solution was dried using a vacuum centrifuge. The peptide pellet was then resuspended in peptide loading buffer (0.1% v/v formic acid, pH 3.5) which was then subjected to peptide identification using a 6560 Ion Mobility LC/Q-TOF (Bruker Daltonics Inc.). Germany). Peptide fingerprint was analyzed using Spectrum Mill software (Release year 2017).