Exosomes from LTBI1
Plasma was harvested and centrifuged at 2,000g at 4 for 30 min to remove cells and cell debris. The supernatant was transferred to a new tube followed by diluting to 20 mL by PBS, and filtered through a filter with a pore size of 0.22 m to exclude particles larger than 200 nm. Then density gradient centrifugation was used to isolate exosomes with 4 ml Tris/sucrose/D2O solution loaded to the bottom of a 70Ti tube, and 20 ml supernatant added to the top of this sucrose cushion, subsequently centrifuging for 75 min at 100,000g at 4 C in a 70Ti rotor. Next, 3.5 mL of the Tris/sucrose/ D2O cushion was removed from the centrifuged tube and transferred to two new centrifuge tubes with 1.75 ml per tube. This mixture was then be diluted with 25 mL PBS, and centrifuged for 70 min at 100,000g at 4 C. Discard the supernatant, the resulting pellet contains the isolated EVs was resuspended in 100 L PBS.