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. 1998 Apr 20;187(8):1261–1271. doi: 10.1084/jem.187.8.1261

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Effect of caspases on PTPC liposomes and isolated mitochondria. (A) Representative DiOC₆(3) fluorescence histograms obtained after treatment of liposomes with various caspases (1.2 U/ml for caspase 1, 10 U/ml for caspase 6) in the presence or absence of the indicated caspase inhibitor (100 μM). (B) Dose dependency of effects obtained with different recombinant caspases on PTPC liposomes. (C) Effect of caspases on the Δψ_m. Mitochondria were treated during 30 min with 5 U caspase/200 μl, followed by determination of the Δψ_m using DiOC₆(3). The protonophore m-chlorophenylhydrazone (50 μM) defined 100% Δψ_m disruption. (D) Release of AIF into the mitochondrial supernatant. Intact mitochondria were treated with the indicated caspase (5 U/200 μl), followed by centrifugation and removal of the supernatant that was tested for apoptogenic activity on isolated HeLa nuclei. The incubation was performed in the presence of tetrapeptide inhibitors (which inhibit caspases but not AIF) or in the presence of Z-VAD.fmk (which inhibits AIF) to exclude that nuclear DNA degradation is a direct caspase effect. Similar results were obtained with mouse and rat (not shown) hepatocyte mitochondria.