Titlebook: Caveolae; Methods and Protocol Cedric M. Blouin Book 2020 Springer Science+Business Media, LLC, part of Springer Nature 2020 electron micro

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https://doi.org/10.1007/978-3-8349-6184-6tients suffering from neuromuscular diseases such as “Caveolinopathies” which are caused by mutations in the . gene encoding for caveolin-3. Human caveolin-3 is a 151 amino acid sized transmembrane protein localized within caveolae, predominantly expressed in cardiac and skeletal muscle cells and in

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Selective Visualization of Caveolae by TEM Using APEX2,, transmission electron microscopy (TEM) has been the method of choice to study caveolae formation and ultrastructure and, more recently, to resolve the sub-caveolar localization of its protein components using novel protein labeling methods for TEM. This chapter describes a protocol for the selecti

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Freeze-Fracture Replica Immunolabeling of Cryopreserved Membrane Compartments, Cultured Cells and Tfunction involving membrane compartments. Freeze-fracturing of biological membranes offers both stunning views onto integral membrane proteins and perpendicular views over wide areas of the membrane at electron microscopical resolution. This information is directly assessable for 3D analyses and qua

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Metamorphosis

Method for Efficient Observation of Caveolin-1 in Plasma Membrane by Microscopy Imaging Analysis,immunofluorescence microscopy, caveolin-1 (CAV1) is visualized as numerous small dots, which are often distributed as a linear array or along the edge of the cell. Although its presence, as well as that of other proteins, can be detected by conventional immunofluorescence microscopy, those results d

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Quantitative Image Analysis of the Spatial Organization and Mobility of Caveolin Aggregates at the omers that further assemble into higher-order structures. Imaging fluorescently labeled caveolin at the plasma membrane with total internal reflection fluorescence (TIRF) microscopy reveals a spatially heterogeneous distribution with aggregates of various sizes. In this chapter, we present a set of

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Spatiotemporal Analysis of Caveolae Dynamics Using Total Internal Reflection Fluorescence Microscopa membrane. Total internal reflection fluorescence microscopy exclusively illuminates molecules in the close vicinity of the glass surface, thereby reducing background fluorescence and enabling observation of the plasma membrane in the glass-attached cells with a high signal-to-noise ratio. Here, we

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Live-Cell FRET Imaging of Phosphorylation-Dependent Caveolin-1 Switch,mple of a noninvasive technique that can be used to achieve this goal at nanometer resolution. FRET-based assays are dependent on the presence of fluorescent probes, such as CFP- and YFP-conjugated protein pairs. Here, we describe an experimental protocol in which live-cell FRET was used to measure

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,Biotin Proximity Labeling to Identify Protein–Protein Interactions for Cavin1,ral cellular environment. Here I describe the use of BioID in HeLa cells to identify proteins that can potentially interact with cavin1, one of the main components of caveolae. Briefly, the method consists in the transfection of the cells with the fusion constructs containing the promiscuous biotin