Skip to main content
. Author manuscript; available in PMC: 2010 Dec 8.
Published in final edited form as: Traffic. 2010 Jan;11(1):110–122. doi: 10.1111/j.1600-0854.2009.01006.x

Figure 6.

Figure 6

Nef increases the production of exosomes in primary T cells. (A) Evaluation of exosome release. Amounts of exosomes released from control primary T cells (lane 1, white bar), those treated with PHA/IL2 (lane 2, grey bar), and those expressing GFP (lane 3, white bar with black stripes) or Nef.GFP (lane 4, black bar) were assessed as in Figs. 1, 3 and 4. Presented are relative protein values in comparison to resting or untransfected cells. Results are presented as the mean ±SD of three independent experiments. 10 μg of these exosomal preparations were analyzed further for the presence of Nef with anti-GFP antibodies by western blotting. Again, capital E and L stand for exosome and cell lysate, respectively. (B) Subcellular localization of Nef. Primary T cells expressing Nef.GFP (green) were stained with anti-CD63 antibodies by indirect immunofluorescence (red). Yellow represents the overlay. White scale bar represents 10 μm. (C) Protein composition of exosomes. Exosomes from primary T cultures treated with PHA/IL2 (lane 2) or those expressing Nef.GFP (lane 4) were analyzed with antibodies directed against conserved exosomal proteins (Hsc70, MHCI, ICAM-1, CD63, CD9, LAMP2, AIP1/Alix, Tsg101, Annexin A2, Lck, AChE, Actin) by western blotting.