Quantification of nonselective bulk autophagy in S. cerevisiae using Pgk1-GFP

Autophagy. 2010 Aug;6(6):794-7. doi: 10.4161/auto.6.6.12348.

Abstract

Rapid estimation of the macroautophagi crate has become of great importance over the past few years. A variety of methods to follow autophagy were established both in S. cerevisiae and the mammalian system. In yeast,measuring the breakdown of GFP-Atg8,and in mammalian cells counting the increase of LC3 puncta, have become the most commonly used assays to quantify autophagy. Here, we provide degradation of Pgk1-GFP followed in immunoblots as a new convenient tool to quantify nonselective bulk autophagy in yeast.

MeSH terms

  • Autophagy*
  • Biological Assay / methods*
  • Blotting, Western
  • Green Fluorescent Proteins / metabolism*
  • Microscopy, Fluorescence
  • Phosphoglycerate Kinase / metabolism*
  • Protein Processing, Post-Translational
  • Recombinant Fusion Proteins / metabolism*
  • Saccharomyces cerevisiae / cytology*
  • Saccharomyces cerevisiae / metabolism*
  • Vacuoles / metabolism

Substances

  • Recombinant Fusion Proteins
  • Green Fluorescent Proteins
  • Phosphoglycerate Kinase