Autolysosomes are organelles that sequester and degrade a portion of the cytoplasm during autophagy. Although autophagosomes are short lived compared to other organelles such as mitochondria, plastids, and peroxisomes, many autolysosomes accumulate in tobacco BY-2 cells cultured under sucrose starvation conditions in the presence of a cysteine protease inhibitor. We here describe our methodology for isolating autolysosomes from BY-2 cells by conventional cell fractionation using a Percoll gradient. The autolysosome fraction separates clearly from fractions containing mitochondria and peroxisomes. It contains acid phosphatase, vacuolar H + -ATPase, and protease activity. Electron micrographs show that the fraction contains partially degraded cytoplasm seen in autolysosomes before isolation although an autolysosome structure is only partially preserved.
CITATION STYLE
Takatsuka, C., Inoue-Aono, Y., & Moriyasu, Y. (2017). Isolation of autolysosomes from tobacco by-2 cells. In Methods in Molecular Biology (Vol. 1511, pp. 151–161). Humana Press Inc. https://doi.org/10.1007/978-1-4939-6533-5_12
Mendeley helps you to discover research relevant for your work.