Exosomes were purified from the conditioned medium (CM) of HepG2 cells using ultracentrifugation methods as described previously.31 Briefly, the CM was collected from HepG2 cells, and then centrifuged at 300 × g for 10 min, 2000 × g for 15 min, and 10,000 × g for 30 min to remove cell debris and additional cellular fragments. After that, the supernatants were ultracentrifuged at 120,000 x g for 70 minutes to isolated exosomes. The pelleted exosomes were then resuspended in 100 μL PBS and stored at −80°C. Nanoparticle tracking analysis (NTA) was used to analyze the quantity and size of exosomes. A JSM-7800F transmission electron microscopy (TEM, StarJoy, Japan) was utilized to identify the morphology of exosomes. The HepG2 cell-derived exosomes were analyzed by Western blotting using the following primary antibodies: anti-TSG101 and anti-CD63.

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