Fig. 5. PKC and Abl mediate ER-stress-induced mitochondria-dependent apoptosis. Neuro2a cells were transfected with control siRNA, or siRNA targeting Abl or PKC. After 48 hours, cells were pretreated with V1-1 (15 minutes, 1 µM) followed by treatment with Tm (5 µg/ml). (A) After 24 hours of Tm treatment, total cell lysates were subjected to western blotting with the indicated antibodies. Data are representative results of three independent experiments. (B) Mitochondrial fractions were isolated from Neuro2a cells treated with Tm for 24 hours after transfection, and subjected to western blot with anti-Bax antibody. TOM20 was used as loading control. Shown are from three independent experiments. (C) TUNEL assay. Neuro2a cells were treated with Tm for 30 hours before staining with TUNEL. Representative data are from two independent experiments. Original magnification was x40.