A number of diseases, including cancer, are linked to perturbation of intracellular signaling pathways.One such pathway is the Ptdlns (3,4,5) P3/Akt (PKB) signaling pathway which is overactivated in a wide range of tumor types.Deactivation of this pathway suppresses the growth of tumor cells and also reduces resistance of tumor cells to chemotherapeutic drug treatment.Akt is a serine/threonine protein kinase with oncogenic and anti-apoptotic activities.Its activation relies on Ptdlns (3,4,5) P3 (an inositol phospholipid localized on the plasma membrane)-mediated translocation of Akt from the cytoplasm to the plasma membrane where Akt gets phosphorylated and activated.Thus, Akt transloeation provides an attractive target for anti-cancer drug discovery.The ultimate goal of this study is to identify Ptdlns (3,4,5) P3/Akt pathway inhibitors that specifically target Akt plasma membrane translocation via conducting a high throughput chemical genetic screening.Recently, we established an experimental system for visualizing this process in live ceils.We used the PH-domain ofAkt (PHAkt) fused with green fluorescent protein (PHAkt-GFP) as a marker for this event and the translocation was elicited by insulin growth factor (IGF).