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Characterization of Oxidative Lipidomics and Autophagy Induction in Chlamydomonas reinhardtii Under Abiotic Stress

Methods Mol Biol. 2021; 
Cristina Ortega-Villasante, Ángel Barón-Sola, Margarita Toledo-Basantes, Flor Martínez, Luis E Hernández
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Catalog Antibody … 14. Anti-ATG8 antibody (AS14 2769, Agrisera) used at a 1:2500 dilution and secondary goat anti-rabbit IgG antibody conjugated with horseradish peroxidase (anti-IgG::HRP, GenScript) used at a 1:4000 dilution. 15 … New Phytol 176:96–107 … Get A Quote

摘要

Autophagy constitutes an essential process triggered by oxidative stress that enables cells to recycle damaged biomolecules and organelles, which is eventually traced by immunodetection with anti-ATG8. In parallel with autophagy induction, carbon metabolism in Chlamydomonas reinhardtii under abiotic stress is diverged toward lipid biosynthesis and lipid droplet accumulation, which can be analyzed by a simple thin-layer chromatography and in vivo staining with the fluorescent probe BODIPY 493/503. We show the responses in Chlamydomonas cells exposed to mercury or cadmium (0-50 μM doses), as examples of oxidative stress-mediated changes in autophagy and lipid metabolism, monitored with the procedures described ... More

关键词

Anti-ATG8 immunoblot, Autophagy, Chlamydomonas reinhardtii, Lipid droplets, Microalga, Oxidative stress, Thin-layer chromatography
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