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Fig. 6. Cav-1 and MAL-GFP are released in prostasomes in PC-3 cells. (A) Whole-mount immunoelectron microscopy of prostasomes isolated from PC-3 cell culture supernatants labelled with anti-CD59 or anti-CD63 mAbs. Bars, 0.2 µm. (B) PC-3 cell-derived prostasomes were analyzed by western blotting using antibodies to CD59 or CD63. The position of molecular weight markers in kDa is indicated on the left. (C) Prostasomes were isolated from the culture supernatant of PC-3 cells stably expressing BENE-myc or MAL-GFP and analyzed by western blotting using antibodies to cav-1 or GFP. The position of the molecular weight markers in kDa is indicated. (D) Prostasomes isolated from PC-3 MAL-GFP cell culture supernatants were subjected to whole-mount immunoelectron microscopy and labelled with anti-GFP antibodies. Bar, 0.2 µm. (E) Prostasomes from PC-3 cells stably expressing BENE-myc were isolated as previously described and then floated on a continuous sucrose density gradient. Aliquots from each fraction (1-12) were run on SDS-PAGE and immunoblotted with anti-cav-1 or anti-CD59 antibodies. Prostasomes floated at a sucrose density of 1.11 g/ml.
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