Fig. 3

EV-mediated ITGA5 transfer from myofibroblasts to HCC cells enhanced the stemness of HCC cells. A Schematic of the in vitro experiment depicting HCC cells (Huh7) cultured with conditioned medium (CM) from myofibroblasts (LX-2 cells with the indicated treatment). B Representative immunofluorescence images of ITGA5 (red) in Huh7 cells treated with or without CM of LX-2 cells (si-Ctrl, si-ITGA5, or treated with GW4869). Scale bars: 20 μm. C Immunoblotting of ITGA5 in Huh7 cells treated with or without CM of LX-2 cells (si-Ctrl or si-ITGA5). D Immunoblotting of ITGA5 in Huh7 cells cultured with or without LX-2 cells (treated with GW4869 or not). E Nanoparticle tracking analysis of LX-2 EVs (left panel); representative transmission electron microscopy image of LX-2 EVs (right panel). Scale bar: 50 nm. F Relative mRNA expression of ITGA5 in Huh7 cells treated with EVs or not. G Immunoblotting of ITGA5 in Huh7 cells treated with 0, 5, or 10 μg/mL EVs for 24 h, or with 10 μg/mL EVs for 0, 24, or 48 h. H Sphere formation assay of Huh7 cells subjected to the indicated treatment. Scale bar: 100 μm. *P < 0.05; **P < 0.01; ***P < 0.001. All data were obtained from three independent experiments per group