Fig. 4

Identification of EVs isolated from the plasma of cervical cancer patients. The EVs were isolated from the fresh peripheral blood of cervical cancer patients using the Total Exosome Isolation Kit. A, a representative transmission electron microscopic image of EVs from plasma of post-irradiated patients was shown. EVs displayed with characteristic bilipid layer and size. B, representative nanoparticle tracking analysis of isolated EVs. C, immunofluorescence staining of characteristic EV marker (CD9) and membrane permeability to CFSE. The fluorescent stained EVs from fetal bovine serum (FBS) were also visualized as the negative control for anti-human CD9. EVs were visualized using a laser-scanning confocal microscope (TCS SP8 STED, Leica). The magnification of image below was 63×10. The total protein of EVs was separated by SDS-PAGE. D, Coomassie Blue staining was performed to demonstrate the significant differences in protein distribution. E, Western-blotting of EV markers (CD9 and TSG101) and non-EV marker (ApoA1) were performed. EV, extracellular vesicle; CFSE, carboxyfluorescein diacetate succinimidyl ester