Fig. 1

ANGPT2 exists on HCC-derived exosomes. a NTA displayed that the majority of isolated exosomes were within 30–150 nm, which is the typical size of exosomes. b Immunoblotting showed the typical exosomal markers (Alix, HSP90, TSG101 and CD63) in isolated exosomes. c Transmission electron microscopic view of isolated exosomes. The isolated exosomes had cup-shaped morphology, were labeled with exosomal marker CD63 (immunogold = 5 nm), and ANGPT2 was also labeled on isolated exosomes by immunogold (immunogold = 10 nm). Scale bar = 100 nm. d IHC demonstrated that the expression of ANGPT2 in HCC tissues (IOD = 270.6 ± 29.36, n = 96) was higher than that in BLD tissues (IOD = 157.3 ± 34.9, n = 11). Scale bar = 100 μm. *P < 0.05, Welch’s t-tests. e Immunoblotting showed that ANGPT2 was positive in exosomes isolated from the sera of both HCC and BLD patients. f ELISA showed that the level of exosomal ANGPT2 isolated from the sera of HCC patients (756.5 ± 20.3 pg/mL, n = 67) was significantly higher than that from the sera of BLD patients (541.3 ± 18.82 pg/mL, n = 26). ***P < 0.001, Welch’s t-tests. g Immunoblotting showed the levels of ANGPT2 in different HCC cell lines and their exosomes (Hep3B, SNU182, SNU387, Li7 and MHCC97H), and the levels of HCC cell-secreted exosomal ANGPT2 were consistent with their corresponding cells. n = 4 for cell groups, n = 5 for exosome groups, **P < 0.01, ***P < 0.001, one-way ANOVA with Tukey’s multiple comparison tests