Autophagy in Development and Disease Methods and Protocols

This detailed book presents state-of-the-art protocols from scientists actively working in autophagy research, a crucial cellular process that regulates numerous cellular functions. The volume explores aspects of autophagy research where a better understanding of its role is vitally important, such as in the development and disease states. 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 Development and Disease: Methods and Protocols serves as an ideal guide for researchers seeking to expand our knowledge of this key cell biological action.

Autophagy Analysis: A Step-by-Step Simple Practical Guide for Immunofluorescence and Western Blotting.- Monitoring Autophagy During Drosophila Oogenesis.- Assessment of Non-Canonical Functions of the Autophagy Proteins in LC3-Associated Phagocytosis (LAP) and LC3-Associated Endocytosis (LANDO).- Monitoring Autophagy with GFP-LC3 Reporter.- Investigating Protein-Protein Interactions of Autophagy-Involved TNIP1.- Proteomic Methods to Study Autophagy in Skin Exposed to Pollutants.- The Chorioallantoic Membrane (CAM) Assay for the Analysis of Starvation-Induced Autophagy.- Keeping an Eye Out for Autophagy in the Cornea: Sample Preparation for Single Cell RNA Sequencing.- Evaluation of Autophagy in Conjunctival Fibroblasts.- Real-Time Visualization of Cholesterol Trafficking in Human Granulosa Cells Using Confocal Live Cell Microscopy as a Tool to Study the Novel Role of Autophagy in Sex Steroid Synthesis.- Visualizing Lipophagy as a New Mechanism of the Synthesis of Sex Steroids in Human Ovary and Testis Using Immunofluorescence Staining Method.- Characterization of Bitter Taste Receptor Dependent Autophagy in Oral Epithelial Cells.- Involvement of Autophagic Machinery in Neuropathogenesis: Targeting and Relevant Methods of Detection.- Assessment of Autophagy in Leishmania Parasites.- Measurement of Autophagic Activity in Cancer Cells with Flow Cytometric Analysis Using Cyto-ID Staining.- Assessing Autophagy Flux in Glioblastoma Temozolomide Resistant Cells.- A Lipidomics Approach to Determine the Role of Lipids and Its Crosstalk with Autophagy in Lung Cancer Metastasis.- Evaluation of Mitochondrial Phagy (Mitophagy) in Human Non-Small Adenocarcinoma Tumor Cells.- Assessment of Stiffness-Dependent Autophagosome Formation and Apoptosis in Embryonal Rhabdomyosarcoma Tumor Cells.- Principles of Hanging Drop Method (Spheroid Formation) in Cell Culture.- Protocol for Testing the Effects of ssDNA Aptamer in HeLa and MCF-7.