Live-cell Imaging of Biosynthetic Protein Transport in Hepatocytes

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Here, we describe a strategy to analyze the exit of apical and basolateral cargo from the trans-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 the ts045 mutant at its non-permissive temperature of 40 °C.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages145-157
Number of pages13
DOIs
StatePublished - 2022

Publication series

NameMethods in Molecular Biology
Volume2544
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • Apical cargo
  • Conditional aggregation domain
  • Hepatocyte
  • Live-cell imaging
  • Protein sorting
  • Protein targeting
  • Protein transport
  • trans-Golgi network

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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