Abstract
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.
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References
Levine B, Kroemer G (2019) Biological functions of autophagy genes: a disease perspective. Cell 176(1–2):11–42
Fernandez AF (2018) Autophagy and proteases: basic study of the autophagic flux by western blot. Methods Mol Biol 1731:73–81
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
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
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
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
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
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
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
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
He C, Levine B (2010) The beclin 1 interactome. Curr Opin Cell Biol 22(2):140–149
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
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
Acknowledgments
S.V.-R. was supported by a German Research Foundation (DFG) grant (VE1153/1-1) and a Josepha und Charlotte von Siebold Habilitandinnen-Förderprogramm (University of Duisburg-Essen). Graphical art in Fig. 2 was partially prepared using Smart Servier Medical Art (https://smart.servier.com/) by Servier (Suresnes, France) licensed under a Creative Commons Attribution 3.0 Unported License (https://creativecommons.org/licenses/by/3.0/).
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Zou, Z., Fernández, Á.F., Jendrossek, V., Vega-Rubín-de-Celis, S. (2022). Studying Autophagy In Vivo in the Mammary Gland and in Xenograft Samples. In: Norberg, H., Norberg, E. (eds) Autophagy and Cancer. Methods in Molecular Biology, vol 2445. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2071-7_16
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DOI: https://doi.org/10.1007/978-1-0716-2071-7_16
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Online ISBN: 978-1-0716-2071-7
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