The uptake assay was performed as previous described [69, 70], the exosomes were labeled with 10 μM PKH67 (a green fluorescent dye, Sigma Aldrich), and incubated in the dark at room temperature for 12 min, and the excess dye was removed using a 100kD ultrafiltration device (Millipore, USA), then the exosomes resuspended in culture medium. Subsequently, the PKH67-labelled exosomes were added to MC3T3-E1 cells and incubated for 24 h at 37◦C. After the cells were fixed with 4% formaldehyde, the nuclei of the cells were stained with 4', 6-diamidino-2-phenylindole (DAPI, Sigma Aldrich), and the cells were visualized by fluorescence under a laser-scanning confocal microscope (LSM710 Carl Zeiss AG).
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