Fig. 6. Cytochrome C release and activation of caspase 9 and 3 by menadione. (A,B) Cytochrome C release in cytosolic (A) and mitochondrial (B) fractions in cell treated with vehicle control, menadione for 2 minutes with or without preincubation with BAPTA AM (n>3). The Y-axis represents the percentage of chemiluminescence of cytochrome C antibodies normalised to maximum fluorescence. Ab and Bb represent western blots of subcellular fractions from control cells, cells treated with BAPTA followed with menadione for 2 minutes and menadione treated cells for 2 minutes, respectively. a.u., arbitrary units. (C) Caspase 9 activity after incubation of cells with menadione for 30 minutes or BAPTA-AM followed by menadione or 100 nM ACh with 100 nM antimycin A or bongkrekic acid followed by menadione. Each column represents the intensity of FAM-LEHD-FMK fluorescence of every propidium-iodide-negative cell in a field of view. (D) Activation of caspase 3 by 30 minutes incubation with menadione, treatment for 30 minutes with 5 pM CCK, 25 nM ACh or bongkrekic acid followed by menadione for 30 minutes or treatment with BAPTA AM followed by menadione application for 30 min. Each column represents the intensity of PhiPhiLux fluorescence of every propidium iodide negative cell in a field of view. a.u., arbitrary units.

Return to article