Fig. 5
Impact of filtration on saliva-derived extracellular vesicles isolated by ultracentrifugation. (A) Representative scheme of the workflow. Briefly, saliva collected from donors was centrifuged successively at 300×g and 3000×g to remove cells and large debris and then small debris respectively. Then, whole saliva supernatant was filtered on 0.45 µm or 0.22 µm filter. The filtrate was subsequently used for EVs isolation as described previously. (B) Concentration of total proteins contained in EVs isolated by ultracentrifugation from 1 mL of human non-filtered saliva (NF) and 0.45 µm or 0.22 µm filtered saliva (n = 3). (C) Quantity of total miRNA contained in EVs isolated by ultracentrifugation from 1 mL of human non-filtered saliva (NF) and 0.45 µm or 0.22 µm filtered saliva (n = 4). (D) miRNA levels in EVs isolated by ultracentrifugation from filtered 0.45 µm or 0.22 µm saliva relative to EVs isolated from non-filtered saliva (NF). Results are given as fold-change versus control non-filtered saliva normalized at 1. (n = 4). *p < 0.05; **p < 0.01.
