Main description:

This volume provides detailed descriptions of prevailing and novel techniques used by experts in the study of PTEN function in disease and biology. The book begins with chapters exploring methods to detect expression levels of PTEN in normal and diseased human specimens; methods to evaluate specific PTEN function in brain cancer; methods that utilize a new biosensor to measure PTEN regulation; and techniques to measure post-transcriptional regulation of PTEN by micoRNAs and ceRNAs. Other chapters present methods describing novel techniques to detect PTEN localization and previously unstudied structural features of PTEN measured through X-Ray Crystallography and Hydrogen Deuterium Exchange Mass Spectrometry. The book concludes with methods to study PTEN function in model organisms including mice and C. elegans. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Authoritative and thorough, PTEN: Methods and Protocols is a valuable collection of methodologies and protocols useful to researchers who are interested in the PTEN field.

Contents:

Part I Introductory Chapters

1. PTEN: History of a Tumor Suppressor
Leonardo Salmena

2. PTEN at 18: Still Growing
Olena Gorbenko and Vuk Stambolic

Part II Methods to Detect PTEN in Disease

3. Immunopathologic Assessment of PTEN Expression
Mireia Castillo-Martin, Tin Htwe Thin, Ana Collazo Lorduy, and Carlos Cordon-Cardo

4. Measurement of PTEN by Flow Cytometry
John F. Woolley and Leonardo Salmena

5. Detecting PTEN and PI3K Signalling in Brain
Guo Zhu and Suzanne J. Baker

6. Germline PTEN Mutation Analysis for PTEN Hamartoma Tumor Syndrome
Joanne Ngeow and Charis Eng

7. Methods for Assessing the In Vivo Role of PTEN in Glucose Homeostasis
Cynthia T. Luk, Stephanie A. Schroer, and Minna Woo

Part III Methods to Study PTEN Regulation
8. Rapid Detection of Dynamic PTEN Regulation in Living Cells Using Intramolecular BRET
Stanislas Misticone, Evelyne Lima-Fernandes, and Mark G.H. Scott

9. Methods for the Identification of PTEN-Targeting MicroRNAs
Andrea Tuccoli, Marianna Vitiello, Andrea Marranci, Francesco Russo, and Laura Poliseno

10.

Post-Transcriptional Regulation of PTEN by Competing Endogenous RNAs
Yvonne Tay and Pier Paolo Pandolfi

11. In Cell and In Vitro Assays to Measure PTEN Ubiquitination
Amit Gupta, Helene Maccario, Nisha Kriplani, and Nicholas R. Leslie

Part IV Methods to Study PTEN Localization
12. Assessing PTEN Subcellular Localization
Anabel Gil, Jose I. Lopez, and Rafael Pulido

13. Methods to Study PTEN in Mitochondria and Endoplasmic Reticulum
Sonia Missiroli, Claudia Morganti, Carlotta Giorgi, and Paolo Pinton

Part IV Methods to Study PTEN Structure

14. Methods in the Study of PTEN Structure: X-Ray Crystallography and Hydrogen Deuterium Exchange Mass Spectrometry
Glenn R. Masson, John E. Burke, and Roger L. Williams

Part VI Methods to Study PTEN and Stem Cell Function

15. Methods for PTEN in Stem Cells and Cancer Stem Cells
Suzanne Schubbert, Jing Jiao, Marcus Ruscetti, Jonathan Nakashima, Shumin Wu, Hong Lei, Qinzhi Xu, Wenkai Yi, Haichuan Zhu, and Hong Wu

Part VII Methods to Study PTEN Using Animal Models

16. Modeling Cancer Associated Mutations of PTEN in Mice
Antonella Papa and Pier Paolo Pandolfi
17. C. elegans Methods to Study PTEN
Shanqing Zheng and Ian D. Chin-Sang