FIG. 3.

Expression of Nef in HIV-1-infected Jurkat cells does not inhibit CD95-, TRAIL-, Ly294002-, or UV light-induced apoptosis. One million Jurkat cells were transduced with nef⁺ or nef-defective HIV-1 NL4-3 particles pseudotyped with the VSV G Env protein. At 3 days posttransduction, the cells were treated with the indicated inducers, and aliquots of the cultures were removed at the indicated time points. Cells were lysed and assayed for caspase-3 and -7 activity (upper panel). GFP expression allowed us to discriminate between uninfected cells and those transduced with the nef⁺ and nef-defective HIV-1 NL4-3 proviral constructs. Apoptosis of GFP-positive cells was assessed by annexin V (AnV, lower panel) staining as described for the bicistronic Nef-GFP expression constructs. All transductions were done in triplicate. Shown are average numbers ± standard deviation. Similar results were obtained in an independent experiment.