Immunofluorescence Analysis by Confocal Microscopy for Detecting Endogenous FOXO

Methods Mol Biol. 2019:1890:143-149. doi: 10.1007/978-1-4939-8900-3_12.

Abstract

Cancer cells are known to inactivate tumor suppressor proteins by triggering their anomalous subcellular location. It has been well established that the aberrant location of FOXO proteins is linked to tumor formation, progression of the same, or resistance to anti-neoplastic treatment. Furthermore, the abnormal location of FOXO has also been considered a potential biomarker for diabetic complications or longevity in different organisms. Here, we describe the immunodetection of endogenous FOXO by confocal microscopy, which can be used as a chemical tool to quantify FOXO expression levels, its cellular location, and even its active/inactive forms with relevant antibodies.

Keywords: Biomarker; Confocal microscopy; FOXO proteins; Immunodetection; Nuclear translocation.

MeSH terms

  • Biomarkers
  • Cell Line, Tumor
  • Cell Nucleus / metabolism
  • Fluorescent Antibody Technique*
  • Forkhead Transcription Factors / genetics
  • Forkhead Transcription Factors / metabolism*
  • Gene Expression
  • Humans
  • Microscopy, Confocal*
  • Protein Transport

Substances

  • Biomarkers
  • Forkhead Transcription Factors