No Nef uptake into 293T by microscopic imaging.

<p>293T (2 x 10<sup>6</sup>) were plated at a 10 cm plate, transfected with GFP, CD81.GFP, or Nef.GFP plasmid, and cultured for 16 hr, followed by direct microscopic imaging with a FITC filter or under the bright field (a 10X objective) (<b>A</b>). Transfected cells were then cultured in exosome-free medium for 3 days. Crude exosomes from the culture medium (40 ml) were prepared as described above, suspended in exosome-free medium, added onto fresh 293T in a polylysine-treated glass bottom dish, and incubated for 3 hr (<b>B</b>) and 12 hr (<b>C</b>). At the end of each incubation, images of the target cells were taken with a FITC filter or under the bright field (a 100X objective). The micrographs were representative of images from multiple fields of two independent experiments.</p>