Fig. 2.

Western blot analysis of cytosolic (A) and mitochondrial (B) cytochrome c from wild-type mice. A, Cytochrome c from the cytosolic fraction in the nonischemic control brain (lane C) and in the ischemic brains (lanes 1–24). Protein (4.1 μg) was loaded per lane. Cytochrome c immunoreactivity was evident as a single band of molecular mass 15 kDa in the cytosolic fraction in the ischemic brain as early as 2 hr after transient FCI (lane 2), whereas it was barely detected in the normal control brain. Cytosolic cytochrome c was sustained until 24 hr after transient FCI. The results of the β-actin analysis are shown in the bottom panel as an internal control. The results shown are representative of three independent studies. B, Western blot analysis of the mitochondrial and cytosolic fractions 4 hr after reperfusion. A significant amount of mitochondrial cytochrome c was detected in the control samples (top lane C under Mitochondria) and was decreased after transient FCI (top lane I underMitochondria). Correspondingly, cytosolic cytochrome c from the same animal showed an increase after transient FCI. Contrarily, COX showed no alteration after FCI (middle panel). Approximately 1.8 μg of protein from the mitochondrial fraction and 6.0 μg of protein from the cytosolic fraction were loaded per lane. C, Nonischemic control;I, ischemic brain 4 hr after reperfusion;CL, control sample of 0.8 μg of mitochondrial protein. A consistent amount of β-actin expression is shown in thebottom lane. The results shown are representative of three independent studies