(A) CX-1 cells were exposed to hyperthermic conditions (42°C) for 1 h in the presence of TRAIL (10-200 ng/ml) and then incubated for 1 h at 37°C in the presence of TRAIL. Equal amounts of protein (20 μg) from cell lysates were separated by SDS-PAGE and immunoblotted with anti-PARP, anti-phosphorylated JNK, anti-JNK, anti-phospho-Bim, or anti-Bim antibody. Actin was used to confirm the equal amount of proteins loaded in each lane. Hyperthermia combined with moderate to high doses of TRAIL (50 – 200ng/ml) shows an increase in activation of the JNK-Bim signaling transduction pathway demonstrating apoptotic cell death. (B, C, D, E, F) Densitometry analysis of the bands from the JNK-Bim signal pathway for PARP (B), p-JNK (C), JNK (D), pBim/Bim (E), and actin (F) was performed.