mTor : methods and protocols / edited by Thomas Weichhart. — New York : Humana Press, c2012. – (58.17/M592/v.821)

Contents

    CONTENTS
    
    Preface
    Contributors
    1 Mammalian Target of Rapamycin: A Signaling Kinase for Every Aspect of Cellular Life
    2 Biochemical and Pharmacological Inhibition of mTOR by Rapamycin and an ATP-Competitive mTOR Inhibitor
    3 Evaluation of the Nutrient-Sensing mTOR Pathway
    4 mTOR Activity Under Hypoxia
    5 Isolation of the mTOR Complexes by Affinity Purification
    6 An In Vitro Assay for the Kinase Activity of mTOR Complex 2
    7 0verexpression or Downregulation of mTOR in Mammalian Cells
    8 Detection of Cytoplasmic and Nuclear Functions of mTOR by Fractionation
    9 Evaluation of Rapamycin-Induced Cell Death 125
    10 Evaluation of mTOR-Regulated mRNA Translation 171
    11 A Genome-wide RNAi Screen for Polypeptides that Alter rpS6 Phosphorylation
    12 Immunohistochemical Analysis of mTOR Activity in Tissues
    13 Assessing Cell Size and Cell Cycle Regulation in Cells with Altered TOR Activity
    14 Quantitative Visualization of Autophagy Induction by mTOR Inhibitors 239
    15 The In Vivo Evaluation of Active-Site TOR Inhibitors in Models of BCR-ABL+ Leukemia 251
    16 Inducible raptor and rictor Knockout Mouse Embryonic Fibroblasts 267
    17 Expanding Human T Regulatory Cells with the mTOR-Inhibitor Rapamycin. 279
    18 Rapamycin-Induced Enhancement of Vaccine Efficacy in Mice 295
    19 Utilizing a Retroviral RNAi System to Investigate In Vivo mTOR Functions in T Cells
    20 Exploring Functional In Vivo Consequences of the Selective Genetic Ablation of mTOR Signaling in T Helper Lymphocytes
    21 Evaluating the Therapeutic Potential of mTOR Inhibitors Using Mouse Genetics
    22 Inhibition of PI3K-Akt-mTOR Signaling in Glioblastoma by mTORC1/2 Inhibitors
    23 Assessing the Function of mTOR in Human Embryonic Stem Cells 361
    24 Video-EEG Monitoring Methods for Characterizing Rodent Models of Tuberous Sclerosis and Epilepsy
    25 A Genetic Model to Dissect the Role of Tsc-mTORC1 in Neuronal Cultures
    26 Tissue-Specific Ablation of Tsc1 in Pancreatic Beta-Cells 407
    27 A Mouse Model of Diet-Induced Obesity and Insulin Resistance 421
    28 Rapamycin as Immunosuppressant in Murine Transplantation Model
    29 Development of ATP-Competitive mTOR Inhibitors
    Index 461