Lipid-Activated Nuclear Receptors

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.

Quantification of Oxysterol Nuclear Receptor Ligands by LC/MS/MS.- A Stable Luciferase Reporter System to Characterise LXR Regulation by Oxysterols and Novel Ligands.- Isolation and Analysis of Human Monocytes and Adipose Tissue Macrophages.- Glucan Encapsulated siRNA Particles (GeRPs) for Specific Gene Silencing in Adipose Tissue Macrophages.- Isolation and Purification of Tissue Resident Macrophages for the Analysis of Nuclear Receptor Activity.- Bone Marrow-Derived Macrophage Immortalization of LXR Nuclear Receptor-Deficient Cells.- Dual Crosslinking Chromatin Immunoprecipitation Protocol for Next Generation Sequencing (ChIPseq) in Macrophages.- Analysis of LXR Nuclear Receptor Cistrome through ChIP-seq Data Bioinformatics.- Quantifying Cellular Cholesterol Efflux.- Methods for Assessing the Effects of LXR Agonists on Macrophage Bacterial Infection.- Measuring Apoptotic Cell Engulfment (Efferocytosis) Efficiency.- Methods to Study Monocyte and MacrophageTrafficking in Atherosclerosis Progression and Resolution.- Preparation of Frozen Liver Tissues for Integrated Omics Analysis.- Identification of Nuclear Receptor Targets By Chromatin Immunoprecipitation in Fatty Liver.- The LPS/D-Galactosamine-Induced Fulminant Hepatitis Model to Assess the Role of Ligand-Activated Nuclear Receptors on the NLRP3 Inflammasome Pathway In Vivo.- Analyzing T-Cell Plasma Membrane Lipids by Flow Cytometry.