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. 2016 Mar 29;7:11150. doi: 10.1038/ncomms11150

Figure 3. Exosomal transfer of miR21 from adipocytes and fibroblasts to ovarian cancer cells.

Figure 3

(a) CAAs and CAFs transiently transfected with FAM-tagged miR21 (miR21-FAM) or without transfection (Control) were co-cultured with mCherry-labelled ovarian cancer SKOV3ip cells for 24 h. Fluorescence microscopy was used to detect the green and red fluorescent signals in SKOV3ip cells. Scale bar, 2 μm. (b) Exosomes were isolated from conditioned media prepared from CAAs and CAFs transfected with FAM-labelled miR21 (miR21-FAM) or without transfection (Control) and added to ovarian cancer SKOV3ip cell cultures. SKOV3ip cells were fixed and the nuclei were stained with 4,6-diamidino-2-phenylindole (DAPI) blue. An SP5 confocal microscope was used to detect the green signals in SKOV3ip cells. Scale bar, 2 μm. (c–h) Red mCherry-labelled ovarian cancer SKOV3ip cells (c) were injected subcutaneously into nude mice, to establish tumours. After 1 week, miR21−/miR21−MEFs transfected with FAM-tagged miR21 (d) were injected intratumorally. After 24 h, tumours were harvested and frozen sections were prepared. A confocal microscopy analysis showed the blue DAPI signals for the nuclei of the two cell types (e), the red mCherry signals for the SKOV3ip cells (f) and the green FAM signals for miR21 (g). Arrowheads indicate the FAM-miR21 signals in the peripheral stromal cells (S) and in some of the red cancer cells (T) (g,h). Scale bar, 5 μm.