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dc.contributor.author
Renna, Felipe Javier
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dc.contributor.author
Herrera Lopez, Malena
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dc.contributor.author
Manifava, Maria
dc.contributor.author
Ktistakis, Nicholas
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dc.contributor.author
Vaccaro, Maria Ines
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dc.date.available
2024-02-22T11:04:50Z
dc.date.issued
2023-04
dc.identifier.citation
Renna, Felipe Javier; Herrera Lopez, Malena; Manifava, Maria; Ktistakis, Nicholas; Vaccaro, Maria Ines; Evaluating Autophagy Levels in Two Different Pancreatic Cell Models Using LC3 Immunofluorescence; Journal of Visualized Experiments; Journal of Visualized Experiments; 194; 4-2023; 1-13
dc.identifier.issn
1940-087X
dc.identifier.uri
http://hdl.handle.net/11336/227937
dc.description.abstract
Autophagy is a specialized catabolic process that selectively degrades cytoplasmic components, including proteins and damaged organelles. Autophagy allows cells to physiologically respond to stress stimuli and, thus, maintain cellular homeostasis. Cancer cells might modulate their autophagy levels to adapt to adverse conditions such as hypoxia, nutrient deficiency, or damage caused by chemotherapy. Ductal pancreatic adenocarcinoma is one of the deadliest types of cancer. Pancreatic cancer cells have high autophagy activity due to the transcriptional upregulation and post-translational activation of autophagy proteins. Here, the PANC-1 cell line was used as a model of pancreatic human cancer cells, and the AR42J pancreatic acinar cell line was used as a physiological model of highly differentiated mammalian cells. This study used the immunofluorescence of microtubule-associated protein light chain 3 (LC3) as an indicator of the status of autophagy activation. LC3 is an autophagy protein that, in basal conditions, shows a diffuse pattern of distribution in the cytoplasm (known as LC3-I in this condition). Autophagy induction triggers the conjugation of LC3 to phosphatidylethanolamine on the surface of newly formed autophagosomes to form LC3-II, a membrane-bound protein that aids in the formation and expansion of autophagosomes. To quantify the number of labeled autophagic structures, the open-source software FIJI was utilized with the aid of the "3D Objects Counter" tool. The measure of the autophagic levels both in physiological conditions and in cancer cells allows us to study the modulation of autophagy under diverse conditions such as hypoxia, chemotherapy treatment, or the knockdown of certain proteins.
dc.format
application/pdf
dc.language.iso
eng
dc.publisher
Journal of Visualized Experiments
dc.rights
info:eu-repo/semantics/openAccess
dc.rights.uri
https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
dc.subject
AUTOPHAGY
dc.subject
LC3
dc.subject
INMUNOFLUORESCENCE
dc.subject
PANCREAS
dc.subject.classification
Métodos de Investigación en Bioquímica
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dc.subject.classification
Ciencias Biológicas
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dc.subject.classification
CIENCIAS NATURALES Y EXACTAS
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dc.title
Evaluating Autophagy Levels in Two Different Pancreatic Cell Models Using LC3 Immunofluorescence
dc.type
info:eu-repo/semantics/article
dc.type
info:ar-repo/semantics/artículo
dc.type
info:eu-repo/semantics/publishedVersion
dc.date.updated
2024-02-20T12:34:13Z
dc.journal.number
194
dc.journal.pagination
1-13
dc.journal.pais
Estados Unidos
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dc.journal.ciudad
Massachussets
dc.description.fil
Fil: Renna, Felipe Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Bioquímica y Medicina Molecular. Universidad de Buenos Aires. Facultad Medicina. Instituto de Bioquímica y Medicina Molecular; Argentina
dc.description.fil
Fil: Herrera Lopez, Malena. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Bioquímica y Medicina Molecular. Universidad de Buenos Aires. Facultad Medicina. Instituto de Bioquímica y Medicina Molecular; Argentina
dc.description.fil
Fil: Manifava, Maria. The Babraham Institute; Reino Unido
dc.description.fil
Fil: Ktistakis, Nicholas. The Babraham Institute; Reino Unido
dc.description.fil
Fil: Vaccaro, Maria Ines. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Bioquímica y Medicina Molecular. Universidad de Buenos Aires. Facultad Medicina. Instituto de Bioquímica y Medicina Molecular; Argentina
dc.journal.title
Journal of Visualized Experiments
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/url/https://www.jove.com/video/65005
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/doi/http://dx.doi.org/10.3791/65005
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