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. 2021 Mar 31;5(7):1238–1251. doi: 10.1002/hep4.1709

FIG. 5.

FIG. 5

EVs collected from CD63‐depleted, HBV‐producing cells possess reduced LHBs protein. (A) Western blot analysis of lysates from HepG2.2.15 cells that were transfected with NT control siRNA or CD63 siRNA. Cells were lysed and EVs were purified by ultracentrifugation at 200,000g for 90 minutes and dissolved in lysis buffer. The expression levels of TSG101, CD63, and LHBs in EVs were determined by western blotting. CD63 depletion resulted in EVs with more TSG101, and less CD63 and LHBs. (B) Corresponding bar graphs quantifying TSG101 and LHBs protein expression from EVs. (C) EVs were isolated and counted by NanoSight with the vesicles ranging in size from 40 nm to 120 nm in diameter. (The y axis shows fold increase.) The numbers of EVs showed no significant difference between EVs with and without CD63 depletion. (D) The DNA titer contained within the EV preparation was determined by RT‐PCR and showed no significant difference between EVs isolated from control cells or cells depleted of CD63 protein. *P < 0.05 and **P < 0.01.