Live-cell Imaging of Biosynthetic Protein Transport in Hepatocytes.
Live-cell Imaging of Biosynthetic Protein Transport in Hepatocytes.
复制标题
肝细胞中生物合成蛋白质运输的活细胞成像。
DOI：
10.1007/978-1-0716-2557-6_10
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发表时间：
2022
期刊：
影响因子：
--
通讯作者：
Müsch,Anne
中科院分区：
文献类型：
--
作者：
Lázaro-Diéguez,Francisco;Müsch,Anne
在这里，我们描述了一种策略，以分析出口的顶端和基底侧货物从thetrans-Golgi网络在原代肝细胞。该方法基于重组腺病毒介导的感染与脉冲追踪方案和荧光蛋白标记的二肽基肽酶IV（DPPIV）和水疱性口炎病毒G（VSVG）货物蛋白的活细胞成像分析的组合，共表达和积累在内质网中通过DPPIV聚集通过工程条件聚集结构域和VSVG通过利用聚集thets 045突变体在其非-允许温度为40 °C。
Here, we describe a strategy to analyze the exit of apical and basolateral cargo from thetrans-Golgi network in primary hepatocytes. The method is based on recombinant adenovirus-mediated infection combined with a pulse-chase regimen and live-cell imaging analysis of fluorescent protein-tagged dipeptidyl peptidase IV (DPPIV) and vesicular stomatitis virus G (VSVG) cargo proteins, coexpressed and accumulated in the endoplasmic reticulum via DPPIV aggregation through an engineered conditional aggregation domain and VSVG by exploiting the aggregation of thets045 mutant at its non-permissive temperature of 40 °C.