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Purification of Specific Cell Population by Fluorescence Activated Cell Sorting FACS
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Method to Purify Phase-Separated p62 Bodies Using Fluorescence-Activated Particle Sorting.

Shuhei Takada1, Masaaki Komatsu2, Hideaki Morishita3,4

  • 1Department of Physiology, Juntendo University Graduate School of Medicine, Bunkyo-ku, Tokyo, Japan.

Methods in Molecular Biology (Clifton, N.J.)
|August 8, 2024
PubMed
Summary

Researchers developed a new method to purify p62 bodies, which are crucial for cellular quality control. This technique aids in discovering new components within these stress-responsive protein aggregates.

Keywords:
AutophagyCell sorterFluorescence-activated particle sortingLiquid–liquid phase separationSelective autophagyUbiquitinated proteinsp62/SQSTM1 body

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Area of Science:

  • Cell Biology
  • Molecular Biology
  • Biochemistry

Background:

  • p62 bodies are ubiquitin-positive cytoplasmic condensates.
  • They form via liquid-liquid phase separation.
  • These bodies are involved in selective autophagy, intracellular quality control, and stress responses.

Purpose of the Study:

  • To describe a method for purifying p62 bodies.
  • To facilitate the identification of novel p62 body constituents.

Main Methods:

  • Purification of p62 bodies using fluorescence-activated particle sorting (FAPS).

Main Results:

  • The described FAPS method enables the isolation of p62 bodies.
  • This purification strategy can be applied under various physiological and stress conditions.

Conclusions:

  • The FAPS method is a valuable tool for studying p62 body composition.
  • This approach will advance our understanding of the roles of p62 bodies in cellular processes.