High-Resolution Imaging of STIM/Orai Subcellular Localization Using Array Confocal Laser Scanning Microscopy.

CRAC CHANNEL: METHODS AND PROTOCOLS(2018)

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摘要
The expression of chimeras that consist of a fluorescent protein (FP) conjugated with a protein of interest provides the ability to visualize, track, and quantify the subcellular localization and dynamics of specific proteins in biological samples. Array confocal laser scanning microscopy is an eminently suitable technique for live-cell imaging of FP-tagged fusion proteins. Here, we describe real-time monitoring of the subcellular dynamics of the stromal-interacting molecule 1 (STIM1) and Orai1, the key protagonists of store-operated Ca entry (SOCE) under resting conditions, and upon Ca mobilization from the endoplasmic reticulum (ER).
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关键词
Ca2+ signaling,Cell transfection,Fluorescence microscopy,Fluorescent proteins,Genetically encoded tools,Image analysis,Live-cell imaging
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