Fig 5. Sucrose density gradient ultracentrifugation in MDM-derived sEVs.

(A) Separation of sEVs by sucrose density gradient. The final sEVs pellet 130,000 x g was placed onto 90–10% sucrose gradient layers, then centrifuged for 200,000 x g for 16h, as indicated. Six fractions were collected from the top to the bottom of the gradient for further WB and confocal experiments. (B) Western blot analysis of sEVs recovered at the fractions F1-F6 (n = 3). 15 μg of total proteins were loaded onto the gel. Relative band intensity was calculated by ImageJ.