Autophagic Flux
Autophagic Flux - Related Videos
09:18
Measuring Diurnal Rhythms in Autophagic and Proteasomal Flux
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Cited by 1 •
2019
We describe our protocol for measuring biological rhythms in protein catabolism via autophagy and the proteasome in mouse liver.
11:39
Assessing Autophagic Flux by Measuring LC3, p62, and LAMP1 Co-localization Using Multispectral Imaging Flow Cytometry
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Cited by 94 •
2017
Here, multispectral imaging flow cytometry with an analytical feature that compares bright detail images of 3 autophagy markers and quantifies their co-localization, along with LC3 spot counting, was used to measure autophagy in an objective, quantitative, and statistically robust manner.
04:33
Quantifying Bulk Autophagic Sequestration Activity in Mammalian Cells
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2025
The video demonstrates an assay for assessing bulk autophagic sequestration activity in mammalian cells. It utilizes electroporation to release cytosolic proteins, including lactate dehydrogenase (LDH). The sequestered LDH and total cellular LDH fractions are determined to assess LDH sequestration, providing insights into the overall bulk sequestration activity.
01:18
Autophagic Cell Death
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2023
Christian de Duve discovered “autophagy,” a process in which cellular components are engulfed by membrane-bound organelles called autophagosomes. The autophagosomes then fuse with lysosomes to digest the enclosed contents. Autophagy is generally activated in cells to prevent cell death. However, cell death is triggered when the damage is beyond repair. Autophagy and Apoptosis Autophagy can activate apoptosis. In normal conditions, the autophagy activating protein Beclin-1 and pro-apoptotic...
01:15
Electric Flux
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2025
The concept of flux describes how much of something goes through a given area. More formally, it is the dot product of a vector field within an area. For a better understanding, consider an open rectangular surface with a small area that is placed in a uniform electric field. The larger the area, the more field lines go through it and, hence, the greater the flux; similarly, the stronger the electric field (represented by a greater density of lines), the greater the flux. On the other hand, if...
FAQs
A single marker measurement cannot show whether the pathway is actively completing degradation or accumulating unfinished intermediates. Autophagic flux adds a time-dependent perspective by tracking material as it proceeds toward lysosomal breakdown. This distinction helps researchers determine whether apparent increases in autophagy reflect stronger pathway activity or a clearance problem that prevents completed processing.
Autophagosomes provide the double-membrane compartment that encloses damaged proteins or organelles, while lysosomes supply an acidic environment and hydrolases, enzymes that digest biological material. Fusion between these compartments connects cargo sequestration with degradation. If either stage is disrupted, cellular material may accumulate, making the completed pathway different from simply forming more autophagosomes.
Researchers compare autophagy markers in samples with and without lysosomal degradation inhibitors. Blocking degradation temporarily reveals how much material would normally be processed through the lysosome, allowing pathway activity to be distinguished from impaired clearance. A marker increase under inhibition therefore provides context for interpreting whether cargo is entering the pathway or failing to move through its final degradative stage.
A typical analysis compares cellular autophagy markers under two conditions: one permitting normal lysosomal degradation and another containing an inhibitor of that degradation. Researchers then evaluate the difference between conditions to infer pathway throughput. This paired design is more informative than examining either condition alone because it connects marker abundance with the fate of cargo inside the pathway.
Nutrient stress can be examined by comparing flux measurements from cells exposed to different nutritional conditions. The resulting differences indicate how cellular quality-control processing changes when resources are limited. Because flux follows cargo from sequestration toward degradation and recycling, the analysis can reveal whether stress is associated with altered pathway activity or reduced completion of lysosomal clearance.
These research areas can involve changes in cellular quality control, including the handling of damaged proteins or organelles. Autophagic flux measurements help determine whether the pathway is responding actively or whether material is accumulating because clearance is impaired. That distinction gives biology researchers a more precise way to interpret pathway behavior across disease-related and age-related cellular contexts.