Autophagy in Differentiation and Tissue Maintenance: Methods and Protocols

Description

This detailed book compiles state-of-the-art protocols from researchers actively working in the area of autophagy, a crucial cellular process that regulates numerous cellular functions. In order to accelerate advances in the field, the volume explores aspects of autophagy research where a better understanding of its role is vitally important, such as in the maintenance of stem cell subpopulations and the regulation of differentiation. Written for the highly successful 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, and tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Autophagy in Differentiation and Tissue Maintenance: Methods and Protocols serves as an ideal guide for researchers seeking to expand our knowledge of this key cell biological action. Front Matter ....Pages i-xii Visualization and Measurement of Multiple Components of the Autophagy Flux (Tracey Evans, Robert Button, Oleg Anichtchik, Shouqing Luo)....Pages 1-12 Mitochondrial Redox Sensor for Drosophila Female Germline Stem Cells (Kiran Suhas Nilangekar, Bhupendra V. Shravage)....Pages 13-20 Induction and Detection of Autophagy in Aged Hematopoietic Stem Cells by Exposing Them to Microvesicles Secreted by HSC-Supportive Mesenchymal Stromal Cells (Rohan S. Kulkarni, Manmohan S. Bajaj, Vaijayanti P. Kale)....Pages 21-34 Skeletal Muscle Lysosomal Function via Cathepsin Activity Measurement (Kristyn Gumpper, Matthew Sermersheim, Michael X. Zhu, Pei-Hui Lin)....Pages 35-43 Autophagy in Adipocyte Differentiation (Zhipeng Tao, Longhua Liu, Louise D. Zheng, Zhiyong Cheng)....Pages 45-53 Determination of Autophagy in the Caco-2 Spontaneously Differentiating Model of Intestinal Epithelial Cells (Sinem Tunçer, Sreeparna Banerjee)....Pages 55-70 Assessing Autophagy in the Leydig Cells (Hui Gao, Chao Liu, Wei Li)....Pages 71-85 Simultaneous Detection of Autophagy and Epithelial to Mesenchymal Transition in the Non-small Cell Lung Cancer Cells (Javad Alizadeh, Shahla Shojaei, Adel Sepanjnia, Mohammad Hashemi, Eftekhar Eftekharpour, Saeid Ghavami)....Pages 87-103 Autophagy in Zebrafish Extraocular Muscle Regeneration (Alfonso Saera-Vila, Phillip E. Kish, Alon Kahana)....Pages 105-117 The Detection Techniques for Autophagy-Associated Cell Death-Related Genes and Proteins: Gene Expression Assay and Immunohistochemistry (Melek Ozturk, Ozlem Dagdeviren Ozsoylemez, Fatma Kaya Dagistanli)....Pages 119-130 Cloning of Autophagy-Related MicroRNAs (Deniz Gulfem Ozturk, Muhammed Kocak, Devrim Gozuacik)....Pages 131-146 Assays to Monitor Aggrephagy in Drosophila Brain (Anne-Claire Jacomin, Ioannis P. Nezis)....Pages 147-157 Methods for Monitoring Autophagy in Silkworm Organs (Aurora Montali, Morena Casartelli, Daniele Bruno, Annalisa Grimaldi, Gianluca Tettamanti)....Pages 159-174 Immunofluorescence Staining Protocols for Major Autophagy Proteins Including LC3, P62, and ULK1 in Mammalian Cells in Response to Normoxia and Hypoxia (Wen Li, Shupeng Li, Yifang Li, Xiaoying Lin, Yongquan Hu, Tian Meng et al.)....Pages 175-185 Identification of Novel Autophagy Inhibitors via Cell-Based High-Content Screening (Georgios Konstantinidis, Sonja Sievers, Yao-Wen Wu)....Pages 187-195 Porcine Cell-Free System to Study Mammalian Sperm Mitophagy (Won-Hee Song, Peter Sutovsky)....Pages 197-207 Monitoring and Measuring Mammalian Autophagy (Sujit K. Bhutia, Prakash P. Praharaj, Chandra S. Bhol, Debasna P. Panigrahi, Kewal K. Mahapatra, Srimanta Patra et al.)....Pages 209-222 Back Matter ....Pages 223-228