Systematic compositional analysis of extracellular vesicles produced by cells undergoing apoptosis, necroptosis or necroptosis

Formation of extracellular vesicles (EVs) has emerged as a novel paradigm in cell-to-cell communication in health and disease. We performed a comparative analysis of steady-state (ssEVs) and cell death-associated EVs (cdEVs) following apoptosis (apoEVs), necroptosis (necEVs), and ferroptosis (ferEVs) using the same cell line. For each EV type, we systematically determined the biophysical and biochemical characteristics, the protein cargo, and the presence of methylated ribosomal RNA. The cell death EVs showed distinctive rRNA methylation patterns compared to ssEVs. RiboMethSeq analysis on EV-extracted RNA for the mapping and quantification of the ribosomal RNA 2'-O-methylations