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Table 4 Uptake inhibition techniques used to evaluate endocytic pathways

From: Extracellular vesicles through the blood–brain barrier: a review

Mechanism

Author

Cells and EVs

Inhibitor (concentration)

Pre-treatment duration

Incubation with EVs

Uptake of Evs

Macropinocytosis

Morad et al

BMECs + 

TdTom-Br-EVs

EIPA (100 μM); cytochalasin D (500 nM)

30 min

3 h

Decreased

Chen et al

BMECs + PKH26-labeled exosomes

EIPA (1 mM); cytochalasin D (20 μM)

30 min

1 h

Decreased

Clathrin-dependent endocytosis

Morad et al

BMECs + 

TdTom-Br-EVs

Chlorpromazine (20 μM); ML141 (100 μM)

30 min

3 h

Decreased

Chen et al

BMECs + PKH26-labeled exosomes

Chlorpromazine (15 μM)

30 min

1 h

Decreased

Lipid raft/caveolae-dependent endocytosis

Morad et al

BMECs + 

TdTom-Br-EVs

Filipin III (10 μM)

30 min

3 h

No effect

Chen et al

BMECs + PKH26-labeled exosomes

Filipin III (5 μM); MβCD (5 mM); nystatin (5 μM)

30 min

1 h

Decreased

  1. EIPA 5-(N-ethyl- N-isopropyl) amiloride, MβCD methyl- β-cyclodextrin