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Full Description
This volume provides an overview of experimental procedures and state-of art methods to investigate FOXOs. Chapters guide readers through biochemical and molecular methods, imaging approaches to monitor the subcellular localization of FOXO factors, omics and bioinformatics approaches, different animal models used in FOXO research, and human studies to investigate the role of FOXO factors in human disease and longevity. 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 FOXO Transcription Factors: Methods and Protocols aims to ensure successful results in the further study of this vital field.
Contents
Introduction to FOXO Biology.- Characterization of microRNAs Regulating FOXO Expression.- Identification of Transcription Factor Binding Sites in the Mouse FoxO1 Promoter.- Investigating Mechanisms that control Ubiquitin-mediated DAF-16/FOXO Protein Turnover.- Phosphorylation of FOXO Proteins as a Key Mechanism to Regulate their Activity.- Methodological Approach for the Evaluation of FOXO as a Positive Regulator of Antioxidant Genes.- Characterization of FOXO Acetylation.- Transcriptional Activity of FoxO Transcription Factors Measured by Luciferase Assays.- Monitoring the Transcriptional Activity of FOXO Transcription Factors by Analyzing their Target Genes.- Using ChIP-based Approaches to Characterize FOXO Recruitment to its Target Promoters.- RNAi Mediated Silencing of FoxO Factors.- Immunofluorescence Analysis by Confocal Microscopy for Detecting Endogenous FOXO.- High Throughput Image-based Screening to Identify Chemical Compounds Capable of Activating FOXO.- Image-based Identification of Chemical Compounds Capable of Trapping FOXO in the Cell Nucleus.- Quantifying Tissue-specific Overexpression of FOXO in Drosophila via mRNA Fluorescence in situ Hybridization using Branched DNA Probe Technology.- Genome-wide Analysis for Identifying FOXO Protein Binding Sites.- Mathematical Modeling of Nuclear Trafficking of FOXO Transcription Factors.- Following Transcriptome to Uncover FOXO Biological Functions.- Hydra as Model to Determine the Role of FoxO in Longevity.- Genetic Ablation of FOXO in Mice to Investigate its Physiological Role.- Analysis of FOXO3 Gene Polymorphisms Associated with Human Longevity.- Analysis of FOXO3a Gene Polymorphism Associated with Asthma.
