Cytokine Effects on Cell Viability and Death of Prostate Carcinoma Cells
Figure 9
Western blot analysis of LNCaP protein expression. (a) and (b) LNCaP cells were treated with TNF-α (10 and 100 ng/mL) for 24 h or IFN-γ (25 ng/mL) for 30, 90 min, and 24 h or pretreated with LY-29004 (20 μM) for 1 h and then treated with IFN-γ (25 ng/mL) for 90 min. Untreated cells for 90 min and 24 h were used as controls (ctrl). (c) and (d) LNCaP cells were treated with IL-1β (5 ng/mL) for 30, 90 min, and 24 h or pretreated with UO126 (30 μM), BAY-117082 (30 μM), or SP600125 (30 μM) for 1 h and then treated with IL-1β (5 ng/mL) for 90 min. Untreated cells for 90 min and 24 h were used as controls (ctrl). (e) and (f) LNCaP and PC-3 cells were treated with IL-1β (5 ng/mL) or TNF-α (10 and 100 ng/mL) or SP600125 (30 μM) or UO126 (30 μM) for 72 h, or pretreated with SP600125 (30 μM) or UO126 (30 μM) for 1 h and then treated with IL-1β (5 ng/mL) for 72 h. Untreated cells for 72 h were used as controls (ctrl). Cell extracts were resolved by SDS-PAGE and analyzed by western blot with antibodies against Bid, Bad, Bax, Bcl-2, Cyclin D1, p-ERK 1/2, p-p38, p-Akt, IκB-α, c-IAP2, and caspase 3. Reprobing with antibody against α-tubulin or β-actin was used as a loading and transfer marker.
