Identification of 24 proteins with conserved extracellular-vesicle-sorting abilities paves the way for EV-based engineering. NBThighlight
EVs for proteomics study were purified using the automated chromatography system . Briefly, EVs were separated on a HiTrap Capto Core 700 column and collected according to the absorbance from a 280 nm UV detector. Samples were further concentrated using 10 kDa cut-off spin-filters and stored at −80 °C before use.Five hundred microliters of concentrated conditioned media was loaded onto SEC columns .
Cs were seeded at 5 × 10cells per well in 24-well plates and incubated overnight. The cells were stimulated with TNF-a for 2 h and treated with mNG-HiBiT-labeled EVs for another 6 h. After trypsinization, cells were resuspended in 100 µL of PBS containing 2% FBS. 4ʹ,6-diamidino-2-phenylindole was added to all samples to exclude dead cells. The samples were measured with MACSQuant Analyzer 10 cytometer .
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