Effects of EAW, luteolin and wedelolactone on t-BHP-induced cytochrome C release and changes in Bcl-2 family proteins. PC12 cells were pretreated with EAW (25 and 50 μg/mL) or luteolin (L10, 10 μM; L20, 20 μM) or wedelolactone (W10, 10 μM; W20, 20 μM) for 24 h, then 100 μM of t-BHP was added for 3 h. (A) Cells were harvested after incubation, and the cytosolic protein extracts were prepared as described in the text and subjected to Western immunoblotting analysis against anti- cytochrome C. The average densitometric value of cytochrome C is shown as the mean ± SD (n = 3). # P < 0.05, compared to the solvent control (0.1% DMSO). *P < 0.05, compared to t-BHP treatment alone. (B) After treatment, the total cell lysates were prepared and subjected to Western immunoblotting analysis against against anti-Bcl-2, -Bcl-xL, -Bax and anti-actin. Actin was used as the loading control. The ratio of Bcl-2/Bax and Bcl-xL/Bax are shown as the mean ± SD (n = 3). # P < 0.05, compared to the solvent control (0.1% DMSO). *P < 0.05, compared to t-BHP treatment alone.