Table 2.

ESCRT-dependent EV biogenesis.

Protein complex	Protein name	General function	EV marker	Biological impact when inhibited	References [#]
ESCRT-0		Cargo sequestration			[55]
	HRS	Binds to and sorts ubiquitinated cargo	ALIX/Flotillin CD63/MHCII/HSC70 or CD81	Reduced intraluminal transport of EGF	[59]
				Decrease in EVs between 50 and 200 nm	[60]
	STAM1	Cargo sorting	CD63/MHCII/HSC70 or CD81	Decrease in EVs less than 50 nm Impaired sorting of MHCII and CD63 in ILV	[60]
ESCRT-I		Bud formation, cargo sequestration			[53,55]
	TSG101	Membrane budding, recognizes ubiquitin	CD63/MHCII/HSC70 or CD81	MHCII and CD63 sorting in ILV is impaired	[60]   [59]
			ALIX/Flotillin	Reduced IL transport of EGF
ESCRT-II		Bud formation, cargo sequestration			[53,55]
	VPS22	Cargo sorting	Syntenin 1/CD63a	Reduced syndecan-CTF and syntenin in exosomes	[61]
ESCRT-III		Vesicle release			[62]
	CHMP4	Membrane scission	Syntenin 1/CD63a	Reduced syndecan-CTF, CD63 and syntenin in exosomes	[61]
Accessory proteins
	VPS4	Dissociates and recycles ESCRT machinery	Syntenin 1/CD63a ALIX/Flotillin	Reduced syntenin and CD63 in exosomes	[61]   [59]
No effect on PLP sorting
	ALIX	Promotes bud formation and abscission	Syntenin 1/CD63a CD63/MHCII/HSC70 or CD81	50% reduction in EVs ranging from 30 to 110 nm Decrease syndecan-CTF, CD63, HSP70 in exosomes MHCII sorting impaired	[61]   [60]
				Decrease syndecan-CTF, CD63, HSP70 in exosomes MHCII sorting impaired

EVs were isolated by differential ultracentrifugation with the 100,000 g pellet containing the EVs that were analyzed unless otherwise indicated.

^aEVs were collected from the 140,000 g pellet.