Fig. 6.
Changes in cytochrome c in cytosolic fraction and mitochondrial membrane potential after treatment with Z-LLLal. Cultured neurons were incubated with 1 μm Z-LLLal for 24 and 48 hr. Then, neurons were lysed in buffer A with 250 mmsucrose by Dounce homogenizer, as described in Materials and Methods. Homogenates were centrifuged and divided into cytosolic and mitochondrial fractions. Cytochrome c was detected in lysates from cytosolic and mitochondrial fractions by Western blot analysis with an anti-cytochrome c monoclonal antibody (PharMingen). Cytosolic fraction of cytochrome c increased, whereas mitochondrial fraction decreased during 48 hr after Z-LLLal treatment. Bcl-x/L expression inhibited cytochrome c release from the mitochondria, whereas DEVD-CHO did not perturb it.