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Related Concept Videos

Autophagy01:27

Autophagy

Autophagy is a self-digesting process by which a cell protects itself from threats both within and outside the cell, ranging from abnormal proteins to invading bacteria. In this process, obsolete components of the cell and invading microbes are degraded by hydrolytic enzymes active in an acidic environment of the lysosomal lumen.
An autophagic pathway consists of a series of signaling events activated in response to diverse stress and physiological conditions such as food deprivation,...

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Related Experiment Video

Updated: Jun 26, 2026

Assessing Autophagic Flux by Measuring LC3, p62, and LAMP1 Co-localization Using Multispectral Imaging Flow Cytometry
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Methodological considerations for assessing autophagy modulators: a study with calcium phosphate precipitates.

Sovan Sarkar1, Viktor Korolchuk, Maurizio Renna

  • 1Department of Medical Genetics, University of Cambridge, Cambridge Institute for Medical Research, Cambridge, UK.

Autophagy
|February 3, 2009
PubMed
Summary

Calcium phosphate precipitates (CPP) initially induce autophagy but later impair autophagosome-lysosome fusion, disrupting autophagy flux in mammalian cells. Researchers recommend caution when using CPP in autophagy studies.

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Last Updated: Jun 26, 2026

Assessing Autophagic Flux by Measuring LC3, p62, and LAMP1 Co-localization Using Multispectral Imaging Flow Cytometry
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Study of Protein-protein Interactions in Autophagy Research
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Study of Protein-protein Interactions in Autophagy Research

Published on: September 9, 2017

Area of Science:

  • Cell Biology
  • Molecular Biology

Background:

  • Autophagy is crucial in physiological and disease states, with modulators explored for therapeutic potential.
  • Mammalian autophagy pathways remain incompletely understood despite yeast studies.
  • Calcium phosphate precipitates (CPP) are used for DNA transfection and were previously reported to induce autophagy.

Purpose of the Study:

  • To investigate the effects of CPP on mammalian autophagy over time.
  • To confirm or refute previous findings on CPP-induced autophagy.
  • To assess the suitability of CPP as a reagent in autophagy research.

Main Methods:

  • Treatment of mammalian cells with CPP.
  • Assaying autophagosome synthesis and autophagy flux at different time points (4h, 6h, 24h).
  • Analysis of autophagy substrates and autophagosome morphology.

Main Results:

  • CPP induced autophagosome synthesis at early time points (4h, 6h).
  • At 24h, CPP treatment reduced autophagy flux by impairing autophagosome-lysosome fusion.
  • Accumulation of autophagy substrates and formation of abnormally large autophagosomes were observed at 24h.

Conclusions:

  • Autophagy modulators should be assayed at multiple time points using diverse methods.
  • CPP exhibits complex effects on autophagy, including impaired fusion at later stages.
  • CPP should be avoided as a transfection reagent in studies focused on autophagy or autophagy-modulated processes like apoptosis.