Main description:

Steroid Receptors: Methods and Protocols presents a selection of techniques that have been recently applied to the analysis of steroid receptors, powerful tools for the advancement of our understanding of both the mechanisms regulating gene transcription and the rapid signaling responses of tissues to signals. Research in this area has generated a wealth of data allowing the elucidation of steroid receptor mechanisms and improving the treatment of many endocrine disorders, above all cancers. Chapters cover methods to analyze gene transcription, chromatin and proteomic modifications, extra-nuclear signaling regulation, development of cell and animal models, and preparation of new antibodies. Written in the 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 protocols and notes on troubleshooting and avoiding known pitfalls.

Authoritative and easily accessible, Steroid Receptors: Methods and Protocols offers an updated view on a variety of modern methods that will hopefully contribute to improving our knowledge on the integration of steroid receptors in single or different functionally connected cellular contexts under a variety of physiological and pathological conditions, above all tumors.

Contents:

1. Progesterone Receptor Interaction with Chromatin

Guillermo P. Vicent, A. Silvina Nacht, Cecilia Ballare, Roser Zaurin, Daniel Soronellas, and Miguel Beato

2. Mapping the Genomic Binding Sites of the Activated Retinoid X Receptor in Murine Bone Marrow-Derived Macrophages using Chromatin Immunoprecipitation Sequencing

Bence Daniel, Balint L. Balint, Zsuzsanna S. Nagy, Laszlo L. Nagy

3. Analysis of Chromatin-Nuclear Receptor Interactions by Laser-Chromatin Immunoprecipitation

Rosaria Benedetti, Mariarosaria Conte, Vincenzo Carafa, Bartolomeo Della Ventura, Carlo Altucci, Raffaele Velotta, Hendrik G. Stunnenberg, Lucia Altucci, and Angela Nebbioso

4. Examining Estrogen Regulation of Cancer Stem Cells through Multicolor Lineage Tracing

Yang Zhang, Gabriel Eades, and Qun Zhou

5. Reporter Mice for the Study of Long Term Effects of Drugs and Toxic Compounds

Nicoletta Rizzi, Balaji Ramachandran, Cristina Vantaggiato, Paolo Ciana, and Adriana Maggi

6. Analysis of Histone post-Translational Modifications in the Control of Chromatin Plasticity Observed at Estrogen-Responsive Sites in Human Breast Cancer Cells

Annalisa Di Santi, Gustavo Cernera, Antimo Migliaccio, and Bruno Perillo

7. In Silico Analysis of Genomic Data for Construction of Nuclear Receptor Network

Yun-Young Park and Ju-Seog Lee

8. Cofactor Profiling of the Glucocorticoid Receptor from a Cellular Environment

Sofie J. Desmet, Lien Dejager, Dorien Clarisse, Jonathan Thommis, Diana Melchers, Niek Bastiaensen, Rob Ruijtenbeek, Ilse M. Beck, Claude Libert, Rene Houtman, Onno C. Meijer, and Karolien De Bosscher

9. Paxillin and Steroid Signaling: from Frog to Human

Stephen R. Hammes, Susanne U. Miedlich, and Aritro Sen

10. Analysis of the Androgen Receptor/Filamin A Complex in Stromal Cells

Pia Giovannelli, Marzia di Donato, Ferdinando Auricchio, and Gabriella Castoria

11. Multi-Well Plate Immunoassays for Measuring Signaling Protein Activations/Deactivations and Membrane Vs. Intracellular Receptor Levels

Cheryl. S. Watson, Yow-Jiun Jeng, Nataliya N. Bulayeva, Celeste C. Finnerty, Luke Y. Koong, Dragoslaca Zivadinovic, Rebecca A. Alyea, Terumi Midoro-Horiuti, Randall M. Goldblum, Noelle. C. Anastasio, Kathryn A. Cunningham, Patricia K. Seitza and Thressa D. Smith

12. Promixity Ligation Assay to Detect and Localize the Interactions of ERa with PI3-K and Src in Breast Cancer Cells and Tumor Samples

Coralie Poulard, Juliette Rambaud, Muriel Le Romancer, and Laura Corbo

13. Phosphoinositide 3-Kinase Assay in Breast Cancer Cell Extracts

Antonio Bilancio and Antimo Migliaccio

14. Rapid Estrogen Effects on Aromatase Phosphorylation in Breast Cancer Cells

Stefania Catalano, Ines Barone, and Sebastiano Ando

15. Mouse Monoclonal Antibodies against Estrogen Receptor

Caterina De Rosa, Valentina Rossi, and Ciro Abbondanza

16. Analysis of Estrogen Receptor Interacting Proteins Using Pull Down Assay and MALDI-MS Methods

Mahendra Kumar Thakur and Vijay Paramanik

17. Analysis of the Conformation of the Androgen Receptor in Spinal Bulbar Muscular Atrophy by Atomic Force Microscopy

Tobias Jochum and Andrew C. B. Cato

18. Imaging of Corticosteroid Receptors in Live Cells

Mayumi Nishi

19. Physiological Techniques in the Study of Rapid Aldosterone Effects

Yamil R. Yusef, Warren Thomas, and Brian J. Harvey

20. Detection of the Glucocorticoid Receptors in Brain Protein Extracts by SDS-PAGE

Fernanda Marques, Joao C. Sousa, Joao J. Cerqueira, and Nuno Sousa