HDAC/HAT Function Assessment and Inhibitor Development

This detailed collection covers how the biological functions of histone deacetylases (HDACs) and histone acetyltransferases (HATs) can be detected in various experimental settings, both in vivo and in vitro. The book also covers the generation and specificity of deacetylase inhibitors and how such agents can be used to test experimental hypotheses. Written for the popular Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, as well as tips on troubleshooting and avoiding known pitfalls.
Comprehensive and practical, HDAC/HAT Function Assessment and Inhibitor Development: Methods and Protocols serves as an ideal guide to this vital area of study.

Includes cutting-edge methods and protocols to study HDACs and HATs Provides step-by-step detail essential for reproducible results Contains key notes and implementation advice from the experts Includes supplementary material: sn.pub/extras

Inhalt

Detection of Autophagy Induction After HDAC Inhibitor Treatment in Leukemic Cells.- Assessment of HDACi-Induced Protein Cleavage by Caspases.- Assessment of HDACi-Induced Cytotoxicity.- Analysis of HDACi-Induced Changes in Chromosomal Passenger Complex Localization.- Analysis of Histone Deacetylase-Dependent Effects on Cell Migration Using the Stripe Assay.- Using Histone Deacetylase Inhibitors to Analyze the Relevance of HDACs for Translation.- Effects of HDACi on Immunological Functions.- DNA Fiber Spreading Assay to Test HDACi Effects on DNA and Its Replication.- Assessing the Effect of Class I Histone Deacetylase Activity on DNA Double-Strand Break Repair by Homologous Recombination.- Establishment and Characterization of Long-Term Cultures Derived from Primary Acute Myeloid Leukemia Cells for HDAC Inhibitor Research.- Assessing the Histone Deacetylase Activity of SIRT6 in Primary Murine Hepatocytes Via Proximity Ligation Assay.- Purification and Analysis of Male Germ Cells from Adult Mouse Testis.- Generation of Tissue-Specific Mouse Models to Analyze HDAC Functions.- Engineering of Conditional Class I Hdac Knockout Mice and Generation of a Time-Spatial Knockout by a Dual Recombination System.- Xenografting of Cancer Cell Lines for In Vivo Screening of the Therapeutic Potential of HDAC Inhibitors.- Generation of Xenotransplants from Human Cancer Biopsies to Assess Anti-Cancer Activities of HDACi.- Phospho-Flow Analysis of Primary Mouse Cells after HDAC Inhibitor Treatment.- Evaluation of Innate Immune Gene Expression Following HDAC Inhibitor Treatment by High Throughput qPCR and PhosFlow Cytometry.- Assessing HDAC Function in the Regulation of Signal Transducer and Activator of Transcription 5 (STAT5) Activity Using Chromatin Immunoprecipitation (ChIP).- Modulation of STAT1-Driven Transcriptional Activity by Histone Deacetylases.- Proximity Ligation Assay to Quantify Foxp3 Acetylation in Regulatory T Cells.- Testing the Effects of SIAH Ubiquitin E3 Ligases on Lysine Acetyl Transferases.- Assessment of HDACi-Induced Acetylation of Non-Histone Proteins by Mass Spectrometry.- Assessing the Role of Paralog-Specific Sumoylation of HDAC1.- Analysis of Histone Deacetylases Sumoylation by Immunoprecipitation Techniques.- How to Distinguish Between the Activity of HDAC1-3 and HDAC6 with Western Blot.- Evaluating the Effect of HDAC8 Inhibition in Malignant Peripheral Nerve Sheath Tumors.- Analyzing the Impact of Pan- and Class-Specific HDACi on Differentiation-Associated Factors.- Encapsulation of the HDACi Ex527 into Liposomes and Polymer-Based Particles.- Preparation of Histone Deacetylase Inhibitor Vorinostat Loaded Poly d, l-lactide-co-glycolide Polymeric Nanoparticles by Nanoprecipitation Method.- Generation and Assessment of Fusions Between HDACi and TKi.