Figure 8

Western blot analysis of cell signaling proteins, listed in methods. PC-3, DU-145 or LNCaP cells were treated either with 1 μM AEE788, 1 mM VPA or 1 nM RAD001, or with the triple drug combination (triple drug). Controls remained untreated. Before evaluation, cells were kept for 2 h in serum-free cell culture medium and subsequently stimulated for 30 min with EGF (100 ng/ml). Cell lysates were then subjected to SDS-PAGE and blotted on the membrane incubated with the respective monoclonal antibodies. β-actin served as the internal control. The figure shows one representative from three separate experiments.