Fig. 1: Highly metastatic MHCC97H-derived exosomes improve migration, chemotaxis and invasion of HLE cells.

a Transmission electron micrograph of MHCC97H exosomes. Arrows indicate exosomes and arrowheads indicate smaller nonexosomal vesicles. Scale bar, 200 nm. b The size distribution of isolated exosomes using Zetasizer Nano ZS90. c Equal amounts of total protein (60 μg) from MHCC97H cells and exosomes were analysed by Western blot for the presence of the exosomal markers CD63, Alix and TSG101, as well as AFP (a recognised HCC marker). The endoplasmatic reticulum protein GRP78 was used as the negative control. d Fluorescence microscopy images of HLE cells treated with fluorescently labelled MHCC97H exosomes (single-stranded RNAs: red; exosomal proteins: green) for 24 h. e Scratch assay for the migration of HLE cells treated with or without MHCC97H-derived exosomes (100 μg/ml). The distance was measured every 6 h for 24 h. f The chemotactic potential of HLE cells treated with or without MHCC97H-derived exosomes (100 μg/ml). The incubation time was 24 h. g Matrigel invasion assay for the invasion of HLE cells treated with or without MHCC97H-derived exosomes (100 μg/ml). The incubation time was 36 h. h Colony formation assay of HLE cells treated with or without MHCC97H-derived exosomes (100 μg/ml). The culture time was 2 weeks. Abbreviation: Exo exosome. *P < 0.05, **P < 0.01 and ***P < 0.001. Scale bar, 1.0 mm. Data are represented as the mean ± S.D. All experiments were repeated at least three times