Titlebook: Autophagy and Cancer; Methods and Protocol Helin Norberg,Erik Norberg Book 2022 The Editor(s) (if applicable) and The Author(s), under excl

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Isolation of Autophagy Competent Lysosomes from Cancer Cells by Differential Large-Scale Multilayerelysosomal content, allowing specific lysosomal investigations induced by autophagy. In this protocol chapter, we describe detailed practical instructions and advices for an efficacious lysosomal enrichment and isolation procedure by differential multilayered density gradient centrifugations using hu

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Modified LC3 Dot Quantification Methoddiscovered for autophagy. At the same time, precise monitoring of autophagy has become important, and western blotting and fluorescence microscopy of the marker protein LC3 is?widely used for this purpose. Here, we describe a modification of the widely used method, number of LC3 dots per cell. This

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A Quantitative Flow Cytometry –Based Method for Autophagy Detection Across the Cell Cycle detection and quantification of various cellular markers in live or fixed cells. Here, a flow cytometry–based assay to characterize autophagy across the cell cycle is described. This method is based on selective plasma membrane permeabilization with digitonin and extraction of membrane-unbound LC3

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Detection of Nuclear Biomarkers for Chromosomal Instabilityenetic material during cell division. Detection of mitotic errors such as misaligned chromosomes or chromosomal bridges (also known as lagging chromosomes) is challenging as it requires the analysis and manual discrimination of chromosomal aberrations in mitotic cells by molecular techniques. In int

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Analysis of Autophagic Vesicles in Mitotic Cellsre commonly used in immunofluorescence and biochemistry assays to evaluate the status of autophagy in adherent cells. During mitosis, cells undergo important morphological changes which alter the position of the central plane, therefore the imaging of dividing cells has to be specifically designed.