Measurement of PTEN Activity in vivo by Imaging Phosphorylated Akt
This chapter describes an indirect approach to measure PTEN’s lipid phosphatase
This chapter describes an indirect approach to measure PTEN’s lipid phosphatase activity in vivo . PTEN counteracts phosphatidylinositol 3-kinase action in dephosphorylating 3-phosphorylated phosphoinositides. Therefore, PtdIns(3,4,5)P3 -dependent activation and phosphorylation of the survival kinase Akt can be used as readout for cellular PTEN activity. Here we have outlined a detailed procedure employing a phosphoserine-specific anti-Akt antibody to examine the content of phosphorylated Akt by immunofluorescence and its dependence on PTEN activity.