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This book covers a wide range of state-of-the-art methodologies and detailed protocols currently used to study the actions that lipid-activated nuclear receptors and their co-regulators have in tissues and immune cell types considered classic metabolic "powerhouses". This includes the liver, adipose tissue, and monocytes/macrophages present in these and other metabolic tissues. While the main focus is on the oxysterol receptor or Liver X Receptor (LXR), the majority of the methods described can be easily applied to multiple nuclear receptors, as well as to other tissues or cell types. 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, Lipid-Activated Nuclear Receptors: Methods and Protocols serves as an ideal guide for researchers pursuing the vital study of nuclear receptor biology and beyond.

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1. Quantification of Oxysterol Nuclear Receptor Ligands by LC/MS/MS

Lilia Magomedova and Carolyn L. Cummins

2. A Stable Luciferase Reporter System to Characterise LXR Regulation by Oxysterols and Novel Ligands

Samantha A. Hutchinson and James L. Thorne

3. Isolation and Analysis of Human Monocytes and Adipose Tissue Macrophages

Jean-Baptiste Julla, Raphaelle Ballaire, Tina Ejlalmanesh, Jean-Fran‡ois Gautier, Nicolas Venteclef, and Fawaz Alzaid

4. Glucan Encapsulated siRNA Particles (GeRPs) for Specific Gene Silencing in Adipose Tissue Macrophages

Emelie Barreby, André Sulen, and Myriam Aouadi

5. Isolation and Purification of Tissue Resident Macrophages for the Analysis of Nuclear Receptor Activity
Laura Alonso-Herranz, Jesús Porcuna, and Mercedes Ricote

6. Bone Marrow-Derived Macrophage Immortalization of LXR Nuclear Receptor-Deficient Cells

Ana Ramón-Vázquez, Juan Vladimir de la Rosa, Carlos Tabraue, and Antonio Castrillo

7. Dual Crosslinking Chromatin-Immunoprecipitation Protocol for Next Generation Sequencing (ChIPseq) in Macrophages

David A. Rollins and Inez Rogatsky

8. Analysis of LXR Nuclear Receptor Cistrome through ChIP-seq Data Bioinformatics

Juan Vladimir de la Rosa, Ana Ramón-Vázquez, Carlos Tabraue, and Antonio Castrillo

9. Quantifying Cellular Cholesterol Efflux

Sabrina Robichaud and Mireille Ouimet

10. Methods for Assessing the Effects of LXR Agonists on Macrophage Bacterial Infection

Estibaliz Glaría, Jonathan Matalonga, and Annabel F. Valledor

11. Measuring Apoptotic Cell Engulfment (Efferocytosis) Efficiency

Matthew C. Gage

12. Methods to Study Monocyte and Macrophage Trafficking in Atherosclerosis Progression and Resolution

Ada Weinstock and Edward A. Fisher

13. Preparation of Frozen Liver Tissues for Integrated Omics Analysis

Ning Liang, Rongrong Fan, Saioa Goni, and Eckardt Treuter

14. Identification of Nuclear Receptor Targets By Chromatin Immunoprecipitation in Fatty Liver

Natalia Becares and Inés Pineda-Torra

15. The LPS/D-Galactosamine-Induced Fulminant Hepatitis Model to Assess the Role of Ligand-Activated Nuclear Receptors on the NLRP3 Inflammasome Pathway In Vivo

Yasmine Sebti, Lise Ferri, Mathilde Zecchin, Justine Beauchamp, Denis Mogilenko, Bart Staels, Hélène Duez, and Benoit Pourcet

16. Analyzing T-Cell Plasma Membrane Lipids by Flow Cytometry

Kirsty E. Waddington, Inés Pineda-Torra, and Elizabeth C. Jury

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