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

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.