Figure 5

Potentiation of cellular response to doxorubicin-induced Akt phosphorylation in MCF7 cells by FAK. MCF7 cells were transiently transfected for 24 hours with a control vector (pcDNA3.1), a focal adhesion kinase (FAK) expression construct or a FAK-related non-kinase (FRNK) expression construct, followed by exposure to 1 μM doxorubicin (Doxo) for a further 24-hour of culture, with or without the addition of the phosphoinositide 3-kinase-specific inhibitor LY294002 (20 μM) 16 hours before the end of exposure to doxorubicin. After treatment the cells were harvested, lysed and subjected to Western blot analyses with antibodies against FAK, FRNK, Ser473-phosphorylated Akt (p-Akt) and total Akt. The densitometric levels of p-Akt were normalized to the respective levels of total Akt in each lane and are shown in the bar graph.