Autophagy is a cellular self-eating process conserved from yeast to humans. Once considered an oddity of yeast cells, autophagy is now known to promote cell survival and regulate processes ranging from immunity and cancer in higher eukaryotes, to development and senescence in both higher and lower eukaryotes. The phenomenon broadly referred to as “autophagy” is composed of two separate processes: macroautophagy and microautophagy. Moreover, autophagy can be used to recycle bulk cytoplasmic contents; to specifically degrade aged, damaged, or excess organelles; or to clear intracellular pathogens. Thus, unraveling the function of autophagy in organismal homeostasis, development, and disease requires a full complement of assays to detect and quantify autophagic initiation and flux. This collection will not only include traditional methods; such as electron microscopy-based detection of autophagosomes, Western blot-based detection of LC3/Atg8 processing and degradation of organelle-specific markers, and fluorescence microscopy-based quantification of LC3/Atg8 puncta, but will also focus on novel strategies that allow detection of autophagy in live cells, quantitative measures of autophagy, and high throughput screening of autophagy-related phenotypes.
Detection of mitochondrial autophagy in adipocyte differentiation
Zhiyong Cheng*1
1University of Florida
WIPI β-propeller proteins as autophagy biomarkers
Tanya Pareek 1,
Tassula Proikas-Cezanne*1
1Institute for Cell Biology, Department of Biology, Faculty of Science, Eberhard Karls Unoiversity Tuebingen, Tuebingen, Germany.