Figure 5

Effects of MP470-Erlotinib combination on phosphorylation of different human RTKs. LNCaP cells were serum starved for 24 hr, pretreated with DMSO (control), Erlotinib (10 μM), MP470 (10 μM), or MP470-Erlotinib (10 μM each) for 2 hr, and then treated with pervanadate (100 μM) for 10 min. For Phosphorylation Antibody Array analysis, 400 μg of cell lysates were incubated with the glass chips at 4°C overnight. After washing for several times, the arrays were incubated with biotin-conjugated anti-Phosphotyrosine for 2 hr, and then with Alexa Fluor 555-conjugated streptavidin for 2 hr. Unbound reagents were removed by washing, and the bound antibodies on the chips were visualized using the GenePix 4000B microarray scanner.