Figure 9

Normal mTOR activity in mTOR+/kd mice. mTOR was immunoprecipitated from heart lysates and the immune complex kinase assays were performed with 4E-BP1 as exogenous substrate, as described in Methods. The degree of phoshorylation of 4E-BP1 and mTOR was visualized via autoradiogram and quantified using a phosphoimager (bottom). Amount of mTOR in immunoprecipitates or in total lysates was assessed by immunoblotting with mTOR antibody. β-tubulin is used as a loading control. Equal amount of 4E-BP1 in the kinase reaction mixture was visualized by Ponceau S staining. Data are representative of two experiments.