Protocol for the isolation of GFP-expressing ferroptosis-dependent extracellular vesicles in in vitrocell culture models

Extracellular vesicles (EVs) are complex structures that transport various DNA, RNA, and protein. Recently, new EV secretion mechanisms have been identified through the iron regulatory system in mammalian cells. We revealed that ferroptosis increases EV secretion, which is named ferroptosis-dependent EVs (FedEVs). Here, we describe a step-by-step procedure to isolate GFP-expressing FedEVs for in vitroanalysis. The FedEVs are further analyzed by imaging and flow cytometry analysis.