From DNA Repair Lab
| Graduate in Biological Sciences (University of São Paulo), develops a PhD project entitled "Autophagy Analysis in response to DNA damage" in the Department of Microbiology, Institute of Biomedical Sciences, University of São Paulo, São Paulo, SP, Brazil.
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Resumo do Projeto
Autophagy is a lysosomal-dependent degradative pathway that promotes recycling of cytoplasmic components, such as organelles and proteins, aiming at maintaining cellular homeostasis.
This process can be activated under different stressful situations, such as starvation, oxidative stress and DNA damage. Deficiencies in this pathway were associated to cardiopathies, cancer, neurodegeneration and aging.
In this lab, we develop a project that aims at investigating autophagy in response to UV-induced lesions on the DNA in skin human fibroblasts, as well as the importance of this degradative pathway in glioma cells exposed to different chemotherapeutic agents.
Autophagosomes in glioma cells (U87MG) stably expressing LC3-GFP (lentiviral transduction) can be analyzed under a fluorescence microscope.
Viable (green cytoplasm; blue and integrate nucleus), necrotic (red nucleus) and early apoptotic cells (green cytoplasm; blue and condensed nucleus) can be analyzed under a fluorescence microscope after stainning with Propidium Iodide, Hoechst and Fluorescein Diacetate.
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