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Related Experiment Video

Updated: Jan 7, 2026

Quantitative Analysis of Autophagy using Advanced 3D Fluorescence Microscopy
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Methods for Measuring TNFα-Induced Autophagy in Cancer Cells.

Sheyda Najafi1, Ehab M Abo-Ali1, Vikas V Dukhande2

  • 1Department of Pharmaceutical Sciences, College of Pharmacy and Health Sciences, St. John's University, Queens, NY, USA.

Methods in Molecular Biology (Clifton, N.J.)
|January 1, 2026
PubMed
Summary
This summary is machine-generated.

Autophagy, a cellular process for homeostasis, can be measured using immunoblotting and fluorescence microscopy. These techniques assess autophagy

Keywords:
ApoptosisAutophagyBafilomycin A1ChloroquineFluorescence microscopyImmunoblottingLC3TNFαp62

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Area of Science:

  • Cellular Biology
  • Molecular Biology

Background:

  • Autophagy is a conserved eukaryotic process vital for cellular homeostasis.
  • It involves recycling damaged organelles and degrading proteins via lysosomes, working with the ubiquitin-proteasome system.
  • Cytokines, like tumor necrosis factor-alpha (TNFα), are signaling proteins that regulate cellular activities, including inflammation and cell death.

Purpose of the Study:

  • To discuss experimental techniques for measuring autophagy.
  • To detail methods for assessing autophagy in response to TNFα treatment.

Main Methods:

  • Immunoblotting
  • Fluorescence microscopy

Main Results:

  • The chapter details how immunoblotting and fluorescence microscopy can quantify autophagy.
  • These methods allow for the measurement of autophagy induction by TNFα.

Conclusions:

  • Experimental techniques like immunoblotting and fluorescence microscopy are effective tools for studying autophagy.
  • These methods enable the investigation of autophagy's role in cellular responses to stimuli such as TNFα.