Kazan Jalal M, Lukacs Gergely L, Apaja Pirjo M, Pause Arnim
Department of Biochemistry, McGill University, Montréal, QC, Canada.
Goodman Cancer Research Centre, McGill University, Montréal, QC, Canada.
Methods Mol Biol. 2019;1998:93-103. doi: 10.1007/978-1-4939-9492-2_7.
The endosomal sorting complexes required for transport (ESCRT) comprise a major trafficking pathway for plasma membrane proteins and are fundamental for ubiquitin-dependent cargo endocytosis. Here, we describe a method for studying the effect of ESCRT complexes on endo-lysosomal membrane trafficking and their role in receptor integrin α5β1 downregulation. Single cell fluorescence ratio image analysis (FRIA), using appropriate fluorescence probes, enables the measurement of dynamics of integrin α5β1 containing vesicles and represents a live cell-based method for studying the role of ESCRTs.
转运所需的内体分选复合体（ESCRT）构成了质膜蛋白的主要运输途径，并且是泛素依赖性货物内吞作用的基础。在此，我们描述了一种研究ESCRT复合体对内溶酶体膜运输的影响及其在受体整合素α5β1下调中的作用的方法。使用适当的荧光探针进行单细胞荧光比率图像分析（FRIA），能够测量含有整合素α5β1的囊泡的动力学，并代表了一种基于活细胞研究ESCRT作用的方法。
