A reporter cell line for real-time imaging of autophagy and apoptosis

Toxicol Lett. 2020 Jun 15:326:23-30. doi: 10.1016/j.toxlet.2020.02.011. Epub 2020 Feb 25.

Abstract

Simultaneous detection of autophagy and apoptosis is important in drug discovery and signaling studies. Here we report, a real-time reporter cell line for the simultaneous detection of apoptosis and autophagy at single-cell level employing stable integration of two fluorescent protein reporters of apoptosis and autophagy. Cells stably expressing EGFP-LC3 fusion was developed initially as a marker for autophagy and subsequently stably expressed with inter-mitochondrial membrane protein SMAC with RFP fusion to detect mitochondrial permeabilization event of apoptosis. The cell lines faithfully reported the LC3 punctae formation and release of intermembrane proteins in response to diverse apoptotic and autophagic stimuli.

Keywords: Apoptosis; Autophagy; Drug screening; Real-time imaging.

MeSH terms

  • Apoptosis / drug effects*
  • Apoptosis / physiology
  • Autophagy / drug effects*
  • Autophagy / physiology
  • Cell Line, Tumor / drug effects*
  • Cell Line, Tumor / physiology
  • Drug Evaluation, Preclinical / methods*
  • Genes, Reporter / drug effects*
  • Genes, Reporter / physiology
  • Green Fluorescent Proteins / drug effects*
  • Green Fluorescent Proteins / physiology
  • HeLa Cells / drug effects*
  • HeLa Cells / physiology
  • Humans

Substances

  • Green Fluorescent Proteins