Figure 2.

Modulation and loading of sEVs with miR-124-3p by transfection with Exo-Fect

(A) sEVs (non-loaded or loaded with miR-124-3p-Cy5) were purified, and the fluorescence of the pellet (sEVs) and supernatant (leftover probe) were quantified. (B) After the loading protocol, the signal intensity in the sEV fraction (pellet) corresponded to over 74% of the total fluorescence observed in the sample. Loading efficiency in (B) was calculated using the following formula: fluorescence intensity of the pellet/(fluorescence intensity of the pellet + fluorescence intensity of the supernatant). (C) qPCR against miR-124-3p in modified sEVs showed an increase in miR-124-3p of over 1,000,000 times the control. (D) sEVs loaded with Exo-Fect and miR-124-3p were treated with RNase and re-purified, and miR-124-3p levels in the vesicles were quantified by qPCR. RNase treatment does not change miR-124-3p levels in sEVs. For all the experiments, n = 3. For the qPCR analysis, U6 was used as housekeeping gene. sEV, small extracellular vesicles; miR-124-3p sEV, miR-124-enriched small extracellular vesicles.