Main description:

This volume presents valuable techniques for studying the class of ligand-activated transcription factors known as nuclear receptors. After a brief overview of the history of the field, chapters cover methods to detect the receptors and their mRNAs in various tissues, protocols to characterize nuclear receptor modulators and activities, their signaling and roles in certain pathogenesis, molecular modeling of nuclear receptor-ligand interactions, as well as the utility of informatics in the field of nuclear receptors. 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 cutting-edge, Nuclear Receptors: Methods and Experimental Protocols seeks to aid researchers working toward furthering our understanding of these vital receptors and their role in numerous pathological conditions.

Contents:

1. Nuclear Receptors: A Historical Perspective

Gisela I. Mazaira, Nadia R. Zgajnar, Cecilia M. Lotufo, Cristina Daneri-Becerra, Jeffrey C. Sivils, Olga B. Soto, Marc B. Cox, and Mario D. Galigniana

2. An Optimized Immunoblotting Protocol for Accurate Detection of Endogenous PGC-1 Isoforms in Various Rodent Tissues

Thomas W. Gettys and Ji Suk Chang

3. Sub-Cellular Localization of NR4A2 Orphan Nuclear Receptor Expression in Human and Mouse Synovial Joint Tissue

Aisling Smyth, Martina Gogarty, Daniel Crean, and Evelyn P. Murphy

4. In Vivo Quantitation of Estrogen Receptor b Subtype Expression in Ovarian Surface Epithelium Using Immunofluorescence Profiling and Confocal Microscopy

Linda S.M. Gulliver

5. In Vivo ChIP-Seq of Nuclear Receptors: A Rough Guide to Transform Frozen Tissues into High Confidence Genome-Wide Binding Profiles

Ashfaq Ali Mir, Kenneth Allen Dyar, Franziska Greulich, Fabiana Quagliarini, Celine Jouffe, Michael Jean Hubert, Marie Charlotte Hemmer, and Nina Henriette Uhlenhaut

6. High-Content Analysis of Constitutive Androstane Receptor Nuclear Translocation

Bryan Mackowiak and Hongbing Wang

7. Antibody Validation Strategy for Nuclear Receptors

Borbala Katona, Ahmed Ibrahim, Marten Sundberg, and Cecilia Williams

8. Immunofluorescence Labeling of Nuclear Receptor Expression in Formalin-Fixed Paraffin-Embedded Tissue

Elizabeth Lightbody and Christopher J.B. Nicol

9. Detection of ADP-Ribosylation of the Androgen Receptor Using the Recombinant Macrodomain AF1521 from Archaeoglobus fulgidus

Teddy Kamata, Chun-Song Yang, Kasey Jividen, Adam Spencer, Natalia Dworak, Luke T. Oostdyk, and Bryce M. Paschal

10. Reconstitution of the Steroid Receptor Heterocomplex

Gisela I. Mazaira and Mario D. Galigniana

11. High-Throughput Imaging of PPIX Using Confocal Microscopy

C. Trent Brewer, Jonathan Low, and Taosheng Chen

12. PGC-1 Overexpression via Local In Vivo Transfection in Mouse Skeletal Muscle

Dongwook Yeo, Chounghun Kang, and Li Li Ji

13. A Reverse Transfection Method for Screening of Nuclear Receptor Activators

Jenni Kublbeck and Paavo Honkakoski

14. Hybrid Reporter Gene Assays: Versatile In Vitro Tools to Characterize Nuclear Receptor Modulators

Jan Heering and Daniel Merk

15. Analysis of the Transcriptional Activity of Retinoic Acid-Related Orphan Receptors (RORs) and Inhibition by Inverse Agonists

Yukimasa Takeda, Hong Soon Kang, and Anton M. Jetten

16. Examining the Role of Nuclear Receptors during In Vivo Chemical-Mediated Breast Tumorigenesis

Jia Yue Shi, Rachel E. Rubino, and Christopher J.B. Nicol

17. Analysis of IL-4/STAT6 Signaling in Macrophages

Syed F. Hassnain Waqas, Grace Ampem, and Tamas Roeszer

18. Isolation and Characterization of Adipose Tissue Macrophages

Grace Ampem and Tamas Roeszer

19. Assessing Mitochondrial Bioenergetics in Isolated Mitochondria from Mouse Heart Tissues Using Oroboros 2k-Oxygraph

Qinqiang Long, Lizhen Huang, Kai Huang, and Qinglin Yang

20. Generation of Skeletal Myocytes from Embryonic Stem Cells through Nuclear Receptor Signaling

Jihong Chen, Hongsheng Liang, Aili Gao, and Qiao Li

21. Obtaining Crystals of PPAR Ligand Binding Domain Bound to Small Molecules

Rebecca L. Frkic and John B. Bruning

22. Molecular Modeling Approach to Study the PPAR -Ligand Interactions

Merilin Al Sharif, Ivanka Tsakovska, Petko Alov, Vessela Vitcheva, Antonia Diukendjieva, and Ilza Pajeva

23. High Dimensional Data Approaches to Understanding Nuclear Hormone Receptor Signaling

Moray J. Campbell