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.
Department Biochemistry II, University Medicine Goettingen, Georg-August University, Goettingen, Germany.
Evelyn
E
Welter
Michael
M
Thumm
Roswitha
R
Krick
D001343:Autophagy; D001681:Biological Assay / Q000379:methods*; D015153:Blotting, Western; D049452:Green Fluorescent Proteins / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D010735:Phosphoglycerate Kinase / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D011993:Recombinant Fusion Proteins / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000166:cytology* / Q000378:metabolism*; D014617:Vacuoles / Q000378:metabolism
D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D010735:Phosphoglycerate Kinase