This second edition volume expands on the previous edition with updates on the latest techniques and technologies used in FOXO research. The chapters in this book are organized into five parts. Part One covers biochemical techniques used to analyze phosphorylation status of FOXO proteins, identify FOXO binding partners, and assess FOXO-specific transcriptional activity. Part Two highlights genetic techniques that are used to identify functional domains and validate disease relevance. Part Three talks about biophysical and bioinformatic methods to study the structure and dynamics of FOXO proteins, and protein-protein interactions. Part Four discusses different fluorescence and imaging techniques to measure FOXO translocation, and Part Five focuses on functional studies in model organisms to characterize cell death regulation and life- and healthspan. 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. Comprehensive and cutting-edge, FOXO Transcription Factors: Methods and Protocols, Second Edition is a valuable resource for both expert and novel researchers who want to learn more about this exciting and developing field of FOXO biology.