Part I: Analysis of Cellular Processes that Are Regulated by HDACs
1. Detection of Autophagy Induction After HDAC Inhibitor Treatment in Leukemic Cells
Anja Goeder, Nisintha Mahendrarajah, and Oliver H. Kramer
2. Assessment of HDACi-Induced Protein Cleavage by Caspases
Fabian Treude, Tobias Gladbach, Jacqueline Plaster, and Joerg Hartkamp
3. Assessment of HDACi-Induced Cytotoxicity
Lisa Blumel, Christian Marx, Marie Kuhne, and Jurgen Sonnemann
4. Analysis of HDACi-Induced Changes in Chromosomal Passenger Complex Localization
Britta Unruhe-Knauf and Shirley K. Knauer
5. Analysis of Histone Deacetylase-Dependent Effects on Cell Migration Using the Stripe Assay
Sonja Mertsch and Solon Thanos
6. Using Histone Deacetylase Inhibitors to Analyze the Relevance of HDACs for Translation Darren M. Hutt, Daniela Martino Roth, Christelle Marchal, and Marion Bouchecareilh
7. Effects of HDACi on Immunological Functions
Rene Winkler and Christian Kosan
8. DNA Fiber Spreading Assay to Test HDACi Effects on DNA and Its Replication
Teodora Nikolova, Anja Goeder, Ann Parplys, and Kerstin Borgmann
9. Assessing the Effect of Class I Histone Deacetylase Activity on DNA Double-Strand Break Repair by Homologous Recombination Andrea Krumm and Wynand P. Roos
Part II: Primary Cell Systems and In Vivo Models to Assess HDAC Functions
10. Establishment and Characterization of Long-Term Cultures Derived from Primary Acute Myeloid Leukemia Cells for HDAC Inhibitor Research
Annette Romanski and Gesine Bug
11. Assessing the Histone Deacetylase Activity of SIRT6 in Primary Murine Hepatocytes Via Proximity Ligation Assay
Katharina Wolf and Susanne Strand
12. Purification and Analysis of Male Germ Cells from Adult Mouse Testis
Thierry Buchou, Minjia Tan, Sophie Barral, Anne-Laure Vitte, Sophie Rousseaux, Juan Arechaga, and Saadi Khochbin
13. Generation of Tissue-Specific Mouse Models to Analyze HDAC Functions
Astrid Hagelkruys, Mirjam A. Moser, and Christian Seiser
14. Engineering of Conditional Class I Hdac Knockout Mice and Generation of a Time-Spatial Knockout by a Dual Recombination System
Sieglinde Bayer and Matthias Wirth
15. Xenografting of Cancer Cell Lines for In Vivo Screening of the Therapeutic Potential of HDAC Inhibitors
Daniel Nettersheim, Sina Jostes, and Hubert Schorle
16. Generation of Xenotransplants from Human Cancer Biopsies to Assess Anti-Cancer Activities of HDACi
Christina Susanne Mullins, Stephanie Bock, Mathias Krohn, and Michael Linnebacher
Part III: Assessment of HDAC-Dependent Effects on Signaling Cascades in Primary and Permanent Cells
17. Phospho-Flow Analysis of Primary Mouse Cells after HDAC Inhibitor Treatment
Dagmar Hildebrand and Katharina F. Kubatzky
18. Evaluation of Innate Immune Gene Expression Following HDAC Inhibitor Treatment by High Throughput qPCR and PhosFlow Cytometry
David Olagnier, Cindy Chiang, and John Hiscott
19. Assessing HDAC Function in the Regulation of Signal Transducer and Activator of Transcription 5 (STAT5) Activity Using Chromatin Immunoprecipitation (ChIP)
Sophia Pinz and Anne Rascle
20. Modulation of STAT1-Driven Transcriptional Activity by Histone Deacetylases
Benjamin Y. Owusu and Lidija Klampfer
21. Proximity Ligation Assay to Quantify Foxp3 Acetylation in Regulatory T Cells
Jing Jiao, Rongxiang Han, Wayne W. Hancock, and Ulf H. Beier
Part IV: PTMs Controlling Acetylation Balances and HDAC Functions
22. Testing the Effects of SIAH Ubiquitin E3 Ligases on Lysine Acetyl Transferases
Jan Hagenbucher, Hilda Stekman, Alfonso Rodriguez-Gil, Michael Kracht, and M. Lienhard Schmitz
23. Assessment of HDACi-Induced Acetylation of Non-Histone Proteins by Mass Spectrometry
Martin Wieczorek, Karl-Heinz Guhrs, and Thorsten Heinzel
24. Assessing the Role of Paralog-Specific Sumoylation of HDAC1
Simona Citro and Susanna Chiocca
25. Analysis of Histone Deacetylases Sumoylation by Immunoprecipitation Techniques
Tobias Wagner, Maren Godmann, and Thorsten Heinzel
Part V: Improved, Novel, and Specific HDACi
26. How to Distinguish Between the Activity of HDAC1-3 and HDAC6 with Western Blot
Mandy Beyer, Nicole Kiweler, Siavosh Mahboobi, and Oliver H. Kramer
27. Evaluating the Effect of HDAC8 Inhibition in Malignant Peripheral Nerve Sheath Tumors
Gonzalo Lopez and Raphael E. Pollock
28. Analyzing the Impact of Pan- and Class-Specific HDACi on Differentiation-Associated Factors
Katrin Noack and Oliver H. Kramer
29. Encapsulation of the HDACi Ex527 into Liposomes and Polymer-Based Particles
Dorle Hennig and Diana Imhof
30. Preparation of Histone Deacetylase Inhibitor Vorinostat Loaded Poly d, l-lactide-co-glycolide Polymeric Nanoparticles by Nanoprecipitation Method
Vilwanathan Ravikumar and Renu Sankar
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31. Generation and Assessment of Fusions Between HDACi and TKi
Siavosh Mahboobi, Bernadette Pilsl, and Andreas Sellmer