PMS1077 Sensitizes TNF-α Induced Apoptosis in Human Prostate Cancer Cells by Blocking NF-κB Signaling Pathway
Figure 5
PMS1077 inhibited TNF-α-induced NF-κB/P65 nuclear translocation.
(A) DU145 cells were pretreated with PMS1077 (50 µM) or TPCA-1(2 µM) for 6 h and followed by TNF-α (20 ng/ml) stimulation for 0.5 h. Cytoplasmic and nuclear extracts representing equal numbers of cells were analyzed by Western blotting using the indicated antibodies. (B) PC-3 cells were pretreated with PMS1077 (50 µM) for 6 h and followed by TNF-α (20 ng/ml) stimulation for 0.5 h. TPCA-1 (2 µM) and DMSO were used as positive NF-κB inhibitor and negative control, respectively. After treatment, cells were stained with primary anti-p65 antibody and Cy3 fluorescein-conjugated secondary antibody (Red), and then the nucleus was counterstained with DAPI (blue) and examined using Fluorescence microscopy. Images were acquired for each fluorescence channel, using suitable filters with 40×objective. The red and blue images were merged using Image J software. All data shown are representative of three independent experiments.