Fig. 6. Disassociation of the PKC-Abl complex by rottlerin reduces ER-stress-induced mitochondrial-dependent apoptosis. Neuro2a cells were treated with Tm (5 µg/ml). After 30 minutes, cells were treated with rottlerin (5 µM). (A) ER and mitochondrial fractions were isolated after a total of 3 hours of Tm treatment. Immunoprecipitates obtained by using anti-Abl antibodies were analyzed by immunoblotting using anti-PKCantibody. Representative data are from three independent experiments. (B) After 24 hours of Tm treatment, mitochondrial and cytosolic fractions were subjected to western blotting using anti-Bax and anti-cytochrome c antibodies, respectively. Tom20 and GAPDH antibodies were used as loading controls for each fraction. Data are representative for two independent experiments. (C) Neuro2a cells were treated with Tm for 30 hours before staining with TUNEL. (Top) Representative data of three independent experiments. Original magnification was x40. (Bottom) TUNEL-positive cells are expressed as a percentage of the number of total cells, as determined by staining after fixation with Hoechst dye. ^*P<0.05 vs Tm treatment.