A Dual HiBiT-GFP-LC3 Lentiviral Reporter for Autophagy Flux Assessment

Methods Mol Biol. 2022:2445:75-98. doi: 10.1007/978-1-0716-2071-7_6.

Abstract

Autophagy is an intracellular degradation process that maintains the cellular homeostasis and it is regulated in multiple ways, both in health and disease. Assessment of autophagic flux in cells is an important approach for understanding the function of autophagy in biological contexts. Here, we describe a new tool for the qualitative and quantitative determination of autophagic flux using a dual lentiviral reporter system that generates a fusion HiBiT-GFP-LC3B protein suitable for generating stable cell lines.

Keywords: Autophagy; HiBiT-GFP-LC3; Lentivirus.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Autophagy* / genetics
  • Cell Line
  • Microtubule-Associated Proteins* / metabolism

Substances

  • Microtubule-Associated Proteins