Fig. 1

Exosomes derived from high metastatic lung cancer cells promotes cell migration and invasion of low metastatic lung cancer cells. A Upper: representative images of wound-healing assay of CL1-0 cells pre-incubated with CL1-0 or CL1-5 conditioned media for 48 h. Scale bar, 200 μm. Lower: the number of migrated cells was calculated. B Upper: representative images of transwell invasion assay of CL1-0 cells pre-incubated with CL1-0 or CL1-5 conditioned media for 48 h. Scale bar, 200 μm. Lower: the number of invaded cells was calculated. C Upper: representative images of wound-healing assay of CL1-0 cells pre-incubated with CL1-0 or CL1-5 exosome-depleted conditioned media for 48 h. Scale bar, 200 μm. Lower: the number of migrated cells was calculated. D Representative transmission electron microscopy images of exosomes derived from CL1-0 and CL1-5 cells. Scale bar, 200 nm. E Nanoparticle tracking analysis of the size distributions and number of exosomes derived from CL1-0 and CL1-5 cells. F Western blot analysis of exosome marker CD9, TSG101 and Flotillin-1 in CL1-0 and CL1-5 cells and exosomes. Calnexin was used as the negative controls. G Immunofluorescence assay of GFP in CL1-0 cells pre-incubated with CD9-GFP containing exosomes derived from CL1-0 or CL1-5 cells. Cells were stained with rhodamine phalloidin and DAPI for F-actin and nuclei, respectively. White arrowheads indicate the CD9-GFP containing exosomes. Scale bar, 20 μm. H Left: representative images of wound-healing assay of CL1-0 cells pre-incubated with CL1-0 or CL1-5 exosomes for 48 h. Scale bar, 200 μm. Right: the number of migrated cells was counted. I Left: representative images of transwell invasion assay of CL1-0 cells pre-incubated with CL1-0 or CL1-5 exosomes for 48 h. Scale bar, 200 μm. Right: the number of invaded cells was counted. Results are presented as mean ± SD from three independent experiments. *P < 0.05. Two-tailed Student’s t-test