Fig. 3
Transfer of miR-7e-5p from tumor cells to macrophages via exosomes. a Scheme showing the transfer of miR-7e-5p-mimics from A20 cells to mouse macrophages. A20 cells were transfected with 50 nM Cy3-miR-7e-5p-mimics (red fluorescence) and seeded in the upper chamber 6 h after transfection. Red fluorescence was detected in macrophages after coculturing with A20 cells for 24 h. Scale bars: 50 μm. b Images from transmission electron microscopy showed bilayer round-shaped particles with diameters ranging from 30 nm to 200 nm collected from the supernatant of A20 cells. Scale bars: 1 μm for lower magnification and 2 μm for higher magnification. c NanoSight particle tracking analysis (NTA) of exosomes revealed that most exosomes had a diameter of approximately 100 to 200 nm. The concentration of vesicles was 2.4 × 108/ml at 150 nm. d Real-time PCR indicated the expression of miR-7e-5p in macrophages after incubation with exosomes extracted from A20 cells after 24 h. A20 cells were previously transfected with miR-7e-5p-mimics (50 nM) or inhibitors (100 nM) for 24 h. Cells treated with transfection reagent only served as a control (mock). Macrophages seeded on six-well plates were incubated with 1.2 × 107 exosomes for 24 h. Statistical test: t-test