FIG. 5.

HIV-1 Nef expressed from the spleen necrosis viral vector, SNV-Nef, induces apoptosis in MVECs in a dose-dependent manner. Representative TUNEL staining of SNV-LacZ and SNV-Nef⁺ vector virion-treated MVECs, infected for 72 h, is shown. The quantities of virions used to infect cells were 10, 100, 500, and 1,000 CFU/ml. As a positive control, MVECs were exposed to DNase at a concentration of 10 μg/ml for 10 min at room temperature. TUNEL assays were performed with the in situ cell death detection kit, TMR Red (Roche Diagnostics). The cells were analyzed by fluorescence microscopy (Olympus System microscope, model BX60, with fluorescence attachment BX-FLA). These results are representative of those from experiments performed in triplicate and repeated at least twice.