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The Detection Techniques for Autophagy-Associated Cell Death-Related Genes and Proteins: Gene Expression Assay and

Melek Ozturk1, Ozlem Dagdeviren Ozsoylemez2, Fatma Kaya Dagistanli3

  • 1Medical Biology Department, Cerrahpasa Faculty of Medicine, Istanbul University, Cerrahpasa, Istanbul, 34098, Turkey. mozturk@istanbul.edu.tr.

Methods in Molecular Biology (Clifton, N.J.)
|September 11, 2017
PubMed

Insights

Autophagy plays a key role in cell survival and homeostasis but can also mediate cell death. This study presents reliable protocols using qRT-PCR and immunohistochemistry to detect autophagy and related cell death markers.

Area of Science:

  • Cell Biology
  • Molecular Biology

Background:

  • Autophagy is crucial for cellular homeostasis and cell survival.
  • Dysregulation of autophagy is linked to diseases like cancer and neurodegeneration.
  • Autophagy can also induce programmed cell death under certain conditions.

Purpose of the Study:

  • To introduce reliable protocols for detecting autophagy and autophagy-associated cell death.
  • To provide methods for analyzing autophagy markers in both cell lines and patient tissues.

Main Methods:

  • Quantitative real-time PCR (qRT-PCR) for gene expression analysis of autophagy markers.
  • Immunohistochemistry for detecting autophagy markers in paraffin-embedded human tissue sections.

Main Results:

  • QRT-PCR offers sensitive, efficient, and automatable gene expression monitoring for autophagy.
  • Immunohistochemistry provides a reliable method for detecting autophagy markers in patient biopsies.

Conclusions:

  • The presented qRT-PCR and immunohistochemistry protocols are effective for studying autophagy and autophagy-associated cell death.
  • These methods facilitate the monitoring of autophagy in cellular and clinical contexts.
Keywords:
AutophagyAutophagy-associated cell deathGene expression assayImmunohistochemistry

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