Kinetics of inhibition of apoptosis by Nfv in Jurkat cells.

(A) Jurkat cells were treated with MeOH (diluent control; ▪), 10 μM AZT (non-PI antiviral control; ■), or 7 μM Nfv (░). Apoptosis was induced with 10 μM CPT every day for 5 days and assessed after annexin-V and propidium iodide staining by flow cytometry. Apoptosis was significantly inhibited by Nfv as early as day 1 (P = .01). (Day 2, P = .04; day 3,P = .005; day 4, P = .001; day 5,P = .03; n = 3.) Similar results were also obtained after stimulation with ActD (10 μg/mL) and CH11 (0.5 μg/mL) (data not shown). (B) Jurkat cells were treated with MeOH (▪) or 7 μM Nfv (░) for 0, 1, 3, 6, or 12 hours and stimulated with 10 μM CPT. Apoptosis was assessed as in (A). Nfv inhibited apoptosis after 1 hour of treatment (P = .01), 3 hours (P = .03), 6 hours (P = .02), and 12 hours (P = .004; n = 3). (C) Dose-response curves. Jurkat cells were treated with increasing doses of MeOH (▪) or Nfv (░) overnight and then stimulated with 10 μM CPT. Apoptosis was assessed as in (A). Whereas 1 μM Nfv did not inhibit apoptosis, 3.5 μM (P = .006; n = 3), 7 μM (P = .02; n = 3), and 10 μM (P = .03; n = 3) all inhibited apoptosis.