Autophagic Lysosomal Inhibition
Autophagic Lysosomal Inhibition - Related Videos
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.
10:16
SorLA and CLC:CLF-1-dependent Downregulation of CNTFRα as Demonstrated by Western Blotting, Inhibition of Lysosomal Enzymes, and Immunocytochemistry
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Cited by 1 •
2017
The present protocol describes how Western blotting and immunocytochemistry combined with inhibition of lysosomal enzymes can be used to demonstrate the downregulation of Ciliary Neurotrophic Factor Receptor-α (CNTFRα), which is conveyed by the interaction between Cytokine-like Factor-1 (CLF-1) and the endocytic receptor sorLA.
03:45
ADI-based Autophagy Induction in Prostate Cancer Cells: An Enzyme-based Technique to Measure Autophagic Response in Cells
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2023
In this video, we demonstrate arginine deiminase-based autophagy induction in prostate cancer cells. Further, advanced fluorescence microscopy can be used to visualize the changes associated with autophagy induction, including the formation and distribution of autophagosomes and lysosomes and their fusion into autolysosomes.
01:31
Lysosomes
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2023
Lysosomes are membrane-enclosed spherical sacs derived from the Golgi apparatus. The most important function of the lysosome is degrading macromolecules and biological polymers that are released during membrane trafficking events such as the secretory, endocytic, autophagic, and phagocytic pathways. The degradation is carried out by several hydrolytic enzymes active in an acidic environment of the lysosomal lumen. These acid hydrolases are involved in cellular processes such as cell signaling,...
01:31
Lysosomes
0 Views •
2023
Lysosomes are membrane-enclosed spherical sacs derived from the Golgi apparatus. The most important function of the lysosome is degrading macromolecules and biological polymers that are released during membrane trafficking events such as the secretory, endocytic, autophagic, and phagocytic pathways. The degradation is carried out by several hydrolytic enzymes active in an acidic environment of the lysosomal lumen. These acid hydrolases are involved in cellular processes such as cell signaling,...
FAQs
Acidification blockade leaves the lysosomal environment unable to support normal degradation, whereas fusion blockade prevents delivery of autophagosomes to lysosomes. Protease inhibition targets breakdown after delivery. These distinct interruption points can produce similar cargo accumulation, so the mechanism of inhibition matters when interpreting where autophagic processing has failed.
LC3-positive structures are useful indicators of accumulated autophagic material, but their increase does not by itself show that autophagy has intensified. A lysosomal block can create the same pattern by stopping downstream clearance. Interpretation therefore requires treating accumulation as evidence of altered flux, rather than as a direct measure of pathway activation.
Comparing lysosomal inhibition with the uninhibited process helps reveal whether cargo normally reaches degradation. If inhibition causes cargo or LC3-positive structures to build up, the difference exposes material that would otherwise be processed. This logic is especially important when testing whether intracellular pathogens are being targeted for xenophagic clearance.
Examine cargo accumulation and LC3-positive structures alongside infection-related outcomes. Changes in microbial survival can indicate whether lysosome-dependent processing contributes to pathogen clearance, while altered antigen processing or inflammatory signaling can reveal broader immune consequences. Together, these observations connect a trafficking defect with host defense rather than treating morphology alone as the outcome.
Apply the intervention while evaluating whether a host-directed treatment changes intracellular microbial survival or infection-associated signaling. If treatment effects differ when lysosomal degradation is interrupted, the comparison can indicate that the treatment depends on, or influences, this arm of host processing. The approach therefore links therapeutic action to a specific cellular degradation stage.
It is most informative when the research question concerns events after autophagic cargo has been formed, including pathogen handling, antigen processing, or inflammatory signaling. By interrupting the lysosome-dependent endpoint, investigators can ask whether observed immune or infection phenotypes require continued degradation. It also helps prevent misreading accumulated structures as proof of heightened autophagy.