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. 2011 Nov 28;287(2):1397–1405. doi: 10.1074/jbc.M111.288662

FIGURE 1.

FIGURE 1.

Suppression of cancerous cell proliferation by exosome isolated from non-cancerous cells. A, cell growth inhibition by a conditioned medium derived from PNT-2 cells is shown. PC-3M-luc cells were incubated for 3 days in a conditioned medium isolated from PC-3M-luc cells, PNT-2 cells, or a culture medium followed by a cell growth assay as described under “Experimental Procedures.” The values on the y axis are depicted relative to the normalized luciferase activity of culture medium-treated cells, which is defined as 1. Each bar is presented as the mean S.E. (n = 3). *, p < 0.05 as compared with culture medium-treated PC-3M-luc cells; Student's t test. B, treatment with GW4869 to donor cells restored the reduced cell growth by the PNT-2-derived CM is shown. Donor PNT-2 cells were incubated in the presence or absence of 10 μm GW4869 for 2 days. The conditioned medium from PC-3M-luc cells was used as a control. The values on the y axis are depicted relative to the normalized luciferase activity of PC-3M-luc-conditioned medium-treated cells, which is defined as 1. Each bar is presented as the mean S.E. (n = 3). *, p < 0.05; Student's t test. C, cell growth inhibition by exosomes derived from PNT-2 cells is shown. PC-3M-luc cells were incubated in exosomes isolated from PNT-2 cells or PC-3M-luc cells followed by a cell growth assay, as described under “Experimental Procedures.” The values on the y axis are depicted relative to the normalized luciferase activity of cells treated with exosomes derived from PC-3M-luc cells is defined as 1. Each bar is presented as the mean S.E. (n = 3). **, p < 0.005, as compared with exosomes isolated from PC-3M-luc cells; Student's t test. D, shown are fluorescent photos of BODIPY-ceramide-labeled PNT-2 and PC-3M-luc cells marked by PKH67. PNT-2 cells and PC-3M-luc cells were labeled with red fluorescent BODIPY-ceramide and green fluorescent PKH67, respectively, as described under “Experimental Procedures.” After treatment of PNT-2 by BODIPY-ceramide, PKH67-labeled PC-3M-luc cells were added. After co-culturing for 3 or 12 h, images were obtained. Fluorescent photos were detected with the Eclipse TE 2000 Inverted Research Microscope, and images were produced using NIS-Elements BR software. Arrowheads show yellow colored cancer cells. The size bar indicates 100 μm.