Novel method for extracting exosomes of hepatocellular carcinoma cells

Figure 3 Exosome protein identification and secretion mechanisms of human liver cancer-associated proteins as analyzed by Western blotting. In total, 30 μg of proteins from Huh-7 whole cell lysates and exosomal proteins were separated by 10% sodium dodecyl sulfate polyacrylamide gel electrophoresis, electrotransferred and incubated with antibodies against exosome protein markers (HSP70 and CD63) and hepatocellular carcinoma-associated proteins (TGM2 and annexin A2). β-actin was used as an internal control. Exosome protein markers (HSP70 and CD63) were identified on the surfaces of exosome membranes. Exosomes secreted by Huh-7 hepatocellular carcinoma cells contained the hepatoma-associated protein TGM2 but were negative for annexin A2. Huh7-exo indicates exosomal proteins isolated from the supernatants of human hepatocellular carcinoma Huh-7 cells; Mr. stands for molecular weight.