microRNA array of LoVo cells and LoVo-Small Extracellular Vesicles

LoVo cells were cultured in sEV-depleted (160,000xg, 16h) complete medium and the supernatant were collected after 72h. sEVs were purified and centrifuged at 100,000xg for 2.5h using a Beckman SW41Ti rotor to know the miRNA in LoVo cells and LoVo-Small Extracellular Vesicles LoVo cells were cultured in sEV-depleted (160,000xg, 16h) complete medium and the supernatant were collected after 72h. After three successive centrifugations at 300×g (5 min), 1,200×g (20 min), and 10,000×g (30 min) at 4℃, sEVs were purified with Amicon Ultra-15 centrifugal filters with a nominal molecular weight limit of 100 kDa and Millex-GP syringe membrane filters with a 0.22-µm pore size (Millipore Express, US) to eliminate debris, then centrifuged at 100,000×g (70 min at 4℃). Vesicles resuspended were loaded onto the top of a step gradient comprising layers of 0.25, 0.5, 1.0, 1.25, and 2M (mol/L) sucrose (S9378, Sigma). The gradients were centrifuged at 100,000xg for 2.5h using a Beckman SW41Ti rotor