Mitotic Exit Methods and Protocols

This volume explores the latest approaches and techniques used to study mitotic exit in diverse model organisms. The chapters in this book cover topics such as transgenic methods generation of RNAi-sensitive cell lines; gene overexpression in heterologue gene expression systems; quantitative live cell imaging and FRET-FLIM; biochemical protocols for analyzing post-translational modifications responsible for mitotic exit regulation; and ways to promote mitosis arrest in disease-associated conditions such as cancer. 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.

Cutting-edge and authoritative, Mitotic Exit: Methods and Protocols is a valuable resource for all researchers interested in learning more about this important and advancing field.

A Transgenic Method to Measure Mitotic Exit in Drosophila Embryos.- Generation of Cdc20 RNAi-Sensitive Cell Lines to Study Mitotic Exit.- Overexpression of GFP Fusions of Regulators of the SAC from Arabidopsis thaliana.- Production of SAC Proteins in a Baculovirus-Based Heterologous Gene Expression System.- Quantitative Live-Cell Imaging to Study Chromatin Segregation and Nuclear Reformation.- Determination of Anaphase Duration by Time-Lapse Microscopy in Budding Yeast.- Visualizing Cartwheel Disassembly Process During Mitosis in Fixed and Live Cells by Fluorescence Microscope.- FRET-FLIM for the Study of Protein-Protein Interactions Underpinning Mitosis Checkpoints.- Using FRET to Define Cdk1-Dependent Ordering of Events during Exit from Second Meiotic M-Phase in Oocytes.- Time-Lapse Imaging to Analyze Cell Fate in Response to Antimitotics.- The Role of Polo-Like Kinase 1 (PLK1) O-GlcNAcylation in Mitosis.- Luciferase-Based Reporter Assay for the Assessment of Aurora A-Kinase Activity in Mitotic Cycle.- Methods for Detecting Phosphorylated Proteins and Observing Intracellular Localization during Mitosis.- Studying Mitotic Phosphorylation in Drosophila.- Inducing Exit from Mitosis by Inactivating Cdk1/Cyclin B in Metaphase-Arrested Cells: A Tool to Study Mitotic Exit and Reestablishment of Interphase.- Targeting Mitotic Exit in Malignant Cells.- The Use of APC/C Antagonists to Promote Mitotic Catastrophe in Cancer Cells.