Fig. 2
Purification and characterization of exosomes from parental and HIV-1 chronically infected U937 cells expressing Nef in a regulatable way. AchE activity and, for HIV-1 infected cells only, HIV-1 Gag CAp24 contents were measured in fractions from 6 to 18 % iodixanol gradients loaded with vesicles obtained by differential centrifugations of supernatants from a U937, b U937 ΔnefHIV-1, c U937 ΔnefHIV-1/wtNef, and d U937 ΔnefHIV-1/NefG2A cells after treatment with HT for 48 h. In the insets, shown are FACS analyses for the presence of both GM1 and CD63 in nanovesicles recovered from pools of AchE positive fractions of iodixanol gradients. Vesicles were bound to aldehyde/sulfate latex beads and then labeled with FITC-conjugated CTX-B and PE-conjugated anti-CD63 mAb. The results are representative of two independent experiments