Studying Autophagy In Vivo in the Mammary Gland and in Xenograft Samples.
Autophagy
Electron microscopy
GFP-LC3
Immunohistochemistry
Immunoprecipitation
Mammary gland
RNAseq
Western blot
Xenografts
Journal
Methods in molecular biology (Clifton, N.J.)
ISSN: 1940-6029
Titre abrégé: Methods Mol Biol
Pays: United States
ID NLM: 9214969
Informations de publication
Date de publication:
2022
2022
Historique:
entrez:
1
1
2022
pubmed:
2
1
2022
medline:
1
4
2022
Statut:
ppublish
Résumé
Autophagy is a dynamic process that can be monitored in multiple ways, both in vitro and in vivo. Studies in mice are a widely used tool to understand multiple diseases and conditions where autophagy plays a role, and therefore autophagic flux measurement in tissues of rodent models are of utmost importance. Here, we present some assays successfully used in determining the autophagy status in the mice mammary gland as well as in xenografts.
Identifiants
pubmed: 34972997
doi: 10.1007/978-1-0716-2071-7_16
doi:
Substances chimiques
Microtubule-Associated Proteins
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
255-272Informations de copyright
© 2022. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.
Références
Levine B, Kroemer G (2019) Biological functions of autophagy genes: a disease perspective. Cell 176(1–2):11–42
doi: 10.1016/j.cell.2018.09.048
Fernandez AF (2018) Autophagy and proteases: basic study of the autophagic flux by western blot. Methods Mol Biol 1731:73–81
doi: 10.1007/978-1-4939-7595-2_8
Mizushima N, Yamamoto A, Matsui M, Yoshimori T, Ohsumi Y (2004) In vivo analysis of autophagy in response to nutrient starvation using transgenic mice expressing a fluorescent autophagosome marker. Mol Biol Cell 15(3):1101–1111
doi: 10.1091/mbc.e03-09-0704
Liang XH, Jackson S, Seaman M, Brown K, Kempkes B, Hibshoosh H et al (1999) Induction of autophagy and inhibition of tumorigenesis by beclin 1. Nature 402(6762):672–676
doi: 10.1038/45257
Qu X, Zou Z, Sun Q, Luby-Phelps K, Cheng P, Hogan RN et al (2007) Autophagy gene-dependent clearance of apoptotic cells during embryonic development. Cell 128(5):931–946
doi: 10.1016/j.cell.2006.12.044
Qu X, Yu J, Bhagat G, Furuya N, Hibshoosh H, Troxel A et al (2003) Promotion of tumorigenesis by heterozygous disruption of the beclin 1 autophagy gene. J Clin Invest 112(12):1809–1820
doi: 10.1172/JCI20039
Yue Z, Jin S, Yang C, Levine AJ, Heintz N (2003) Beclin 1, an autophagy gene essential for early embryonic development, is a haploinsufficient tumor suppressor. Proc Natl Acad Sci U S A 100(25):15077–15082
doi: 10.1073/pnas.2436255100
Fernandez AF, Sebti S, Wei Y, Zou Z, Shi M, McMillan KL et al (2018) Disruption of the beclin 1-BCL2 autophagy regulatory complex promotes longevity in mice. Nature 558(7708):136–140
doi: 10.1038/s41586-018-0162-7
Vega-Rubin-de-Celis S, Zou Z, Fernandez AF, Ci B, Kim M, Xiao G et al (2018) Increased autophagy blocks HER2-mediated breast tumorigenesis. Proc Natl Acad Sci U S A 115(16):4176–4181
doi: 10.1073/pnas.1717800115
Vega-Rubin-de-Celis S (2019) The role of beclin 1-dependent autophagy in cancer. Biology (Basel) 9(1):4
Vega-Rubin-de-Celis S, Kinch L, Pena-Llopis S (2020) Regulation of beclin 1-mediated autophagy by oncogenic tyrosine kinases. Int J Mol Sci 21(23):9210
doi: 10.3390/ijms21239210
He C, Levine B (2010) The beclin 1 interactome. Curr Opin Cell Biol 22(2):140–149
doi: 10.1016/j.ceb.2010.01.001
Wei Y, Zou Z, Becker N, Anderson M, Sumpter R, Xiao G et al (2013) EGFR-mediated Beclin 1 phosphorylation in autophagy suppression, tumor progression, and tumor chemoresistance. Cell 154(6):1269–1284
doi: 10.1016/j.cell.2013.08.015
Shoji-Kawata S, Sumpter R, Leveno M, Campbell GR, Zou Z, Kinch L et al (2013) Identification of a candidate therapeutic autophagy-inducing peptide. Nature 494(7436):201–206
doi: 10.1038/nature11866