Zinc Protoporphyrin Upregulates Heme Oxygenase-1 in PC-3 Cells via the Stress Response Pathway
Figure 6
Western blot analysis of HO-1 protein induced by ZnPP in PC-3 in the presence of various kinase inhibitors or antioxidant. (a) PC-3 cells were pretreated with nothing (lane 2), 5 μM LY294002 (lane 3), 10 μM U0126 (lane 4), 5 μM Ro 31-8220 (lane 5), 2 μM rottlerin (lane 6), or 5 μM Ro 32-0432 (lane 7) for 2 h, and then treated with 10 μM ZnPP for 24 h. Control cells were treated with equal amount of DMSO for 24 h (lane 1). Total proteins (50 μg) were analyzed by western blot as described in Section 2, using specific antibodies against β-actin and HO-1. Immunoreactive protein bands detected by WesternDot 625 appeared as fluorescent bands. (b) PC-3 cells were pretreated with 5 μM myristoylated PKC-θ pseudosubstrate (lane 1), 5 μM myristoylated PKC-ζ pseudosubstrate (lane 2), 5 μM myristoylated PKC-η pseudosubstrate (lane 3), 5 μM PKC-β inhibitor (lane 4), or nothing (lane 5) for 2 h, and then treated with 10 μM ZnPP for 24 h. Control cells were treated with equal amount of DMSO for 24 h (lane 6). Western blot analysis was done as described above.
