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. 2005 Nov 7;171(3):493–503. doi: 10.1083/jcb.200507069

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Copyright © 2005, The Rockefeller University Press

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Figure 2. — Fusion activity is associated with exosomes. (A) Fusogenic activity is enriched by sequential centrifugation. Conditioned medium from IE^H cells (S0) was sequentially centrifuged at 500, 16,000, 70,000, and 110,000 g. A pellet from each centrifugation was reconstituted and used in the fusion assay with I0 cells. (B) P70 obtained as in A was treated with trypsin or buffer (see Materials and methods), and the fusogenicity of the resulting preparations was determined as in A. (C) Electron micrograph of P70 from IE^H cells. (D and E) P70 was fractionated by floating in a sucrose gradient, and the fractions were analyzed for CD81 (D) and in the fusion assay with I0 cells (E). (F) Exosomes from I0 cells are not fusogenic. Indicated concentrations of exosomes from either I0 or IE^H cells were tested in the fusion assay with I0 cells. The experiments in A, B, and F were performed at least three times. The data in D and E are from one of three experiments, and the data in A and B are from the same experiment. The error bars in A, B, and F indicate SDs.