Nfv inhibits loss of mitochondrial membrane potential (Δψm) as well as cytochrome-c release and caspase-8 cleavage.
(A) Jurkat cells were treated overnight with 10 μM CPT. Δψm of cells in the (i) dead (annexin negative propidium iodide positive), (ii) healthy (annexin negative propidium iodide negative), (iii) apoptotic (annexin positive propidium iodide negative), and (iv) early dead (annexin positive propidium iodide positive) cell populations were simultaneously assessed using DiOC6. (B) Cumulative data from (A) refer to the percentage of cells exhibiting a DiOC6-low phenotype. Data are representative of 3 independent experiments. pi indicates propidium iodide. (C) Nfv treatment inhibits cytochrome-c release. Jurkat cells were treated overnight with MeOH or 7 μM Nfv, and apoptosis was induced by stimulation with 10 μM CPT or 0.5 μg/mL CH11 for 8 hours, as indicated. Immunoblots were then performed of Hsp 70 in the mitochondrial pellet (upper) and cytosol (middle), and of cytochrome-c release in cytosol (lower; 40μg of protein per lane). Blots are representative of 2 independent experiments.
