Fig. 1.

Extracellular vesicles purified from DKs-8, DLD-1, and DKO-1 cells have exosomal characteristics. A, exosomes were purified from conditioned medium of DKs-8, DLD-1, and DKO-1 cells via differential centrifugation as described in the text. One hundred micrograms of whole cell lysate (WCL) protein and 10 μg of exosome (EXO) protein were resolved via SDS-PAGE and Western blotted with the indicated antibodies. Experiments were performed at least in triplicate. B, two independent exosome preparations from DKs-8 and DKO-1 cells were negatively stained and viewed via TEM. Representative field images show vesicles with a smooth, saucer-like morphology characteristic of exosomes (left-hand panels). The mean diameter of 200 vesicles was calculated from TEM images, and the exosome frequency was plotted for indicated size groups of 10 nm each (right-hand panels). The mean diameter for DKs-8 is 59.2 nm ± 14.2 nm, and the mean diameter of DKO-1 is 56.3 nm ± 17.9 nm. Scale bar represents 100 nm.