Fig. 4.

JNK activation in response to blockage of NF-κB due to induction of GADD45α and γ. (A) JNK kinase assays after transfection with GADD45α, β, and γ. DU145 cells were transfected with GADD45α, β, and γ expression vectors (1 μg) and analyzed by using the SAPK/JNK assay kit (Cell Signaling Technology). (B) DU145 cells were infected with AdCMVIκBα or AdCMVβ-gal and protein extracts were collected 48 h after infection. The proteins were analyzed by Western blots (Top and Middle) by using anti-MKK4 antibody (Cell Signaling Technology), anti-phospho-JNK (Cell Signaling Technology), and anti-total JNK (Cell Signaling Technology), and c-jun phosphorylation (Bottom) showed MKK4 and JNK activity in prostate cancer cells at 24, 48, and 72 h after infection with each virus.