Department of Pharmacology and Toxicology, Medical College of Georgia, Augusta University, Augusta, GA 30912, USA.
STAR Protoc. 2024 Jun 21;5(2):102955. doi: 10.1016/j.xpro.2024.102955. Epub 2024 Mar 13.
Here, we present a protocol for visualization and quantification of the recruitment of newly synthesized G protein-coupled receptors (GPCRs) to coat protein complex II vesicles and GPCR transport from the endoplasmic reticulum through the Golgi to the cell surface in the retention using the selective hooks assay. We describe steps for plasmid construction, cell transfection, transport synchronization, confocal microscope imaging, and quantification. This protocol is also applicable for studying the transport of non-GPCR cargoes. For complete details on the use and execution of this protocol, please refer to Xu et al..
在此,我们提供了一种使用选择性钩附实验检测新合成的 G 蛋白偶联受体 (GPCR) 募集到被膜蛋白复合物 II 小泡和 GPCR 从内质网通过高尔基体到细胞表面的保留中的运输的可视化和定量方法的实验方案。我们描述了质粒构建、细胞转染、运输同步、共聚焦显微镜成像和定量的步骤。该方案也适用于研究非 GPCR 货物的运输。有关此方案的使用和执行的完整详细信息,请参阅 Xu 等人的文章。