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Isolation of Nucleoli for Characterization of Nucleolar Contents to Uncover Clues to Metastatic Progression

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Metastasis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2294))

Abstract

Nucleolar isolation is a crucial technique for the study of nucleolar contents and regulation of ribosome biogenesis. Lysed cells are spun through various concentrations of sucrose and magnesium chloride to separate the notoriously dense nucleoli from the rest of the cell. Here we describe isolation of nucleoli from the breast cancer cell line MDA-MB-468. The resulting nucleolar fraction is subjected to immunoblotting to confirm the purity of the nucleolar fraction.

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References

  1. Weeks SE, Metge BJ, Samant RS (2019) The nucleolus: a central response hub for the stressors that drive cancer progression. Cell Mol Life Sci 76(22):4511–4524. https://doi.org/10.1007/s00018-019-03231-0

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Ruggero D (2012) Revisiting the nucleolus: from marker to dynamic integrator of cancer signaling. Sci Signal 5(241):pe38. https://doi.org/10.1126/scisignal.2003477

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Derenzini M, Montanaro L, Trere D (2009) What the nucleolus says to a tumour pathologist. Histopathology 54(6):753–762. https://doi.org/10.1111/j.1365-2559.2008.03168.x

    Article  PubMed  Google Scholar 

  4. Prakash V, Carson BB, Feenstra JM, Dass RA, Sekyrova P, Hoshino A, Petersen J, Guo Y, Parks MM, Kurylo CM, Batchelder JE, Haller K, Hashimoto A, Rundqivst H, Condeelis JS, Allis CD, Drygin D, Nieto MA, Andang M, Percipalle P, Bergh J, Adameyko I, Farrants AO, Hartman J, Lyden D, Pietras K, Blanchard SC, Vincent CT (2019) Ribosome biogenesis during cell cycle arrest fuels EMT in development and disease. Nat Commun 10(1):2110. https://doi.org/10.1038/s41467-019-10100-8

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Busch H, Muramatsu M, Adams H, Steele WJ, Liau MC, Smetana K (1963) Isolation of nucleoli. Exp Cell Res 24(SUPPL9):150–163

    Article  Google Scholar 

  6. Hacot S, Coute Y, Belin S, Albaret MA, Mertani HC, Sanchez JC, Rosa-Calatrava M, Diaz JJ (2010) Isolation of nucleoli. Curr Protoc Cell Biol. Chapter 3:Unit3 36. https://doi.org/10.1002/0471143030.cb0336s47

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Correspondence to Rajeev S. Samant .

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Weeks, S.E., Samant, R.S. (2021). Isolation of Nucleoli for Characterization of Nucleolar Contents to Uncover Clues to Metastatic Progression. In: Stein, U.S. (eds) Metastasis. Methods in Molecular Biology, vol 2294. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1350-4_19

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  • DOI: https://doi.org/10.1007/978-1-0716-1350-4_19

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1349-8

  • Online ISBN: 978-1-0716-1350-4

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