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Effect of GW4869 treatment on exosomes secretion

VC Vincent Clément
VR Vincent Roy
BP Bastien Paré
CG Cassandra R. Goulet
LD Lydia Touzel Deschênes
FB François Berthod
SB Stéphane Bolduc
FG François Gros-Louis
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Fibroblasts were seeded at a density of 3 × 104 cells/cm2 in 12-well plates in complete DMEM. After 48 h, cells were pre-treated with 30 μM GW4869 (Sigma-Aldrich) or DMSO for 48 h. For exosomes production, fibroblasts were then incubated with complete DMEM without phenol red (Wisent Bioproducts) supplemented with 5% of bovine exosome-depleted FBS, antibiotics and 30 μM GW4869 (Sigma-Aldrich) or DMSO, in 8% CO2 at 37 °C for 72 h. Exosomes were isolated with Total Exosome Isolation Reagent from cell culture media (Invitrogen) as described above. Finally, total exosomal proteins was quantified using a standard Bradford assay (Bio-Rad).

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