FIGURE 3.

Artocarpin induces apoptosis via the mitochondrial pathway. (A) Cells were treated with various artocarpin concentrations for 24 h. Effect of artocarpin on U87 mitochondrial depolarization was measured with JC-1 staining and a fluorescence plate reader. (B) Cells were treated with artocarpin for 24 h then the Bad, Bax, and Bcl-2 protein expression levels were determined by western blot. (C) Cells were treated with artocarpin for 16- or 24 h. Cytosolic- and mitochondrial fractions were prepared and subjected to western blot with anti-cytochrome c antibody. GAPDH was used as a marker protein for cytosolic fractions. COX IV was used as a marker protein for mitochondrial fractions. (D) Cells were pretreated with MCI-186 (10 μM) for 1 h then treated with artocarpin for 24 h. Bad, Bax, and Bcl-2 protein expression levels were determined by western blot. (E) Cells were pretreated with MCI-186 (10 μM) for 1 h then treated with artocarpin for 24 h. Cytosolic- and mitochondrial fractions were prepared and subjected to western blot with anti-cytochrome c antibody. Data are expressed as means ± SE of three independent experiments. ^#P < 0.01 compared with cells exposed to vehicle.