Lanzuolo Chiara, Marasca Federica (eds.) Polycomb Group Proteins: Methods and Protocols

2nd edition. — Humana Press, 2023. — 258 p. — (Methods in Molecular Biology 2655). — ISBN 978-1-0716-3142-3.This volume explores the latest technologies used in the Polycomb Group of proteins field and helps scientists--working on PcG proteins--investigate all functions of PcG proteins in diverse cellular contexts. The chapters in this book cover topics such the distribution of histone marks by CUT&Tag in Drosophila embryos; Co-IP in mammalian cells; replication timing of gene loci in different cell cycle phases; STORM and electronmicroscopy and relative data analysis; and polycomb mediated epigenetic modification in spheroids. 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 comprehensive, Polycomb Group Proteins: Methods and Protocols, Second Edition is a valuable tool for all researchers looking to expand their knowledge of this developing field.Analyzing the Genome-Wide Distribution of Histone Marks by CUT&​Tag in Drosophila Embryos
Chromatin Preparation and Chromatin Immunoprecipitation from Drosophila Heads
Detecting Cell Compartment-Specific PRC2-RNA Interactions via UV-RIP
Chromosome Conformation Capture Followed by Genome-Wide Sequencing (Hi-C) in Drosophila Embryos
Physics-Based Polymer Models to Probe Chromosome Structure in Single Molecules
Co-Immunoprecipitat​ion (Co-Ip) in Mammalian Cells
Site-Directed Mutagenesis Protocol to Determine the Role of Amino Acid Residues in Polycomb Group (PcG) Protein Function
Isolation of Chromatin Proteins by Genome Capture
Dynamic Interactome of PRC2-EZH1 Complex Using Tandem-Affinity Purification and Quantitative Mass Spectrometry
Replication Timing of Gene Loci in Different Cell Cycle Phases
Polycomb Bodies Detection in Murine Fibromuscular Stroma from Skin, Skeletal Muscles, and Aortic Tissues
Segmentation, 3D Reconstruction, and Analysis of PcG Proteins in Fluorescence Microscopy Images in Different Cell Culture Conditions
STORM Microscopy and Cluster Analysis for PcG Studies
Quantitative Analysis of PcG-Associated Condensates by Stochastic Optical Reconstruction Microscopy (STORM)
Immunoelectron Microscopy Methods
Differentiation of hPSCs to Study PRC2 Role in Cell-Fate Specification and Neurodevelopment​
Analysis of Polycomb Epigenetic Marks in HeLa Spheroids
Establishment and Maintenance of Human CRC-Derived Organoids for PcG Studies