Proximity Ligation in Situ Assay to Monitor Autophagy-Related Protein Interactions and Autophagy in Cancer Cells

Methods Mol Biol. 2022:2543:167-178. doi: 10.1007/978-1-0716-2553-8_14.

Abstract

Proximity ligation in situ assay (PLISA) is a powerful method to quantify endogen protein-protein interactions in cells and simultaneously identify localization of these interactions. PLISA can be used to quantify autophagy flux and can as well be adapted to assess global autophagy (SQSTM1/P62-LC3B interaction) or specific autophagy, such as mitophagy (NIX-LC3B). Here, we describe a step-by-step method to monitor autophagy using PLISA in adherent cancer cells.

Keywords: Amplification; Autophagy; Confocal microscopy; LC3B; Ligation; PLISA.

MeSH terms

  • Autophagy*
  • Autophagy-Related Proteins / metabolism
  • Mitophagy
  • Neoplasms*
  • Sequestosome-1 Protein / metabolism

Substances

  • Autophagy-Related Proteins
  • Sequestosome-1 Protein