Live-cell Imaging of Biosynthetic Protein Transport in Hepatocytes

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.

Keywords: Apical cargo; Conditional aggregation domain; Hepatocyte; Live-cell imaging; Protein sorting; Protein targeting; Protein transport; trans-Golgi network.