Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

Export of Misfolded Proteins out of the ER01:32

Export of Misfolded Proteins out of the ER

3.7K
After folding, the ER assesses the quality of secretory and membrane proteins. The correctly folded proteins are cleared by the calnexin cycle for transport to their final destination, while misfolded proteins are held back in the ER lumen. The ER chaperones attempt to unfold and refold the misfolded proteins but sometimes fail to achieve the correct native conformation. Such terminally misfolded proteins are then exported to the cytosol by ER-associated degradation or ERAD pathway for...
3.7K
Protein Import into the Peroxisomes01:27

Protein Import into the Peroxisomes

3.6K
Cells contain membrane-bound organelles called peroxisomes that oxidize organic molecules by transferring hydrogen atoms to oxygen, producing hydrogen peroxide. Peroxisomes enzymatically convert the released hydrogen peroxide into water and oxygen.
Peroxisomal Protein Import:
Peroxisomes lack the genetic machinery required to code for their own proteins. Hence, most peroxisomal membrane, lumenal and transmembrane proteins are synthesized in the cytoplasm or ER and transported to the peroxisome...
3.6K
ER Retrieval Pathway01:45

ER Retrieval Pathway

3.9K
In the secretory pathway, vesicles transport proteins from one cellular compartment to another in forward transport to deliver the protein to its correct location. Occasionally, misfolded proteins and incorrect proteins escape their original compartments, and a retrieval pathway is used to return the escaped proteins to their original compartment.
The ER uses many checkpoints to prevent the entry of incorrectly folded or a resident protein as cargo onto a transport vesicle. These mechanisms...
3.9K
Delivery Pathways to the Lysosome01:36

Delivery Pathways to the Lysosome

6.6K
Eukaryotic cells use different mechanisms to eliminate toxic waste obsolete and worn-out substances. Lysosomes play a pivotal role in this, and hence, these substances are carried to the lysosome from other parts of the cell and extracellular space through different pathways. The most elaborately studied pathways to the lysosome are the endocytic pathways.
Endocytosis
In endocytosis, the cell membrane takes up macromolecules and particles from the surrounding medium. Clathrin-mediated...
6.6K
Autophagy01:27

Autophagy

4.4K
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,...
4.4K

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Quantifying the biomechanical relationship between labial augmentation prosthesis thickness and upper lip displacement: A cross-sectional clinical study.

The Journal of prosthetic dentistry·2026
Same author

Graded activation of the CLEAR network through mTORC1 suppression on cargo-harboring lysosomes.

Cell reports·2026
Same author

Extracellular release in the quality control of the mammalian mitochondria.

Journal of biomedical science·2023
Same author

Nanoscale details of mitochondrial constriction revealed by cryoelectron tomography.

Biophysical journal·2023
Same author

Golgi quality control and autophagy.

IUBMB life·2022
Same author

The correlation study on homocysteine, blood lipids and blood glucose levels in patients with cerebral infarction.

American journal of translational research·2021

Related Experiment Video

Updated: Jul 28, 2025

Monitoring Stub1-Mediated Pexophagy
08:26

Monitoring Stub1-Mediated Pexophagy

Published on: May 12, 2023

1.6K

Monitoring Stub1-Mediated Pexophagy.

Bo-Hua Chen1, Wei Yuan Yang2

  • 1Institute of Biological Chemistry, Academia Sinica.

Journal of Visualized Experiments : Jove
|May 29, 2023
PubMed
Summary

Mammalian cells remove peroxisomes using Stub1-mediated pexophagy. This process can be triggered and monitored using dyes that generate reactive oxygen species (ROS), offering control over peroxisome quantity and quality.

More Related Videos

Analysis of Autophagy in Penicillium chrysogenum by Using Starvation Pads in Combination With Fluorescence Microscopy
06:51

Analysis of Autophagy in Penicillium chrysogenum by Using Starvation Pads in Combination With Fluorescence Microscopy

Published on: February 1, 2015

8.6K
In Vitro and In Vivo Detection of Mitophagy in Human Cells, C. Elegans, and Mice
08:40

In Vitro and In Vivo Detection of Mitophagy in Human Cells, C. Elegans, and Mice

Published on: November 22, 2017

17.6K

Related Experiment Videos

Last Updated: Jul 28, 2025

Monitoring Stub1-Mediated Pexophagy
08:26

Monitoring Stub1-Mediated Pexophagy

Published on: May 12, 2023

1.6K
Analysis of Autophagy in Penicillium chrysogenum by Using Starvation Pads in Combination With Fluorescence Microscopy
06:51

Analysis of Autophagy in Penicillium chrysogenum by Using Starvation Pads in Combination With Fluorescence Microscopy

Published on: February 1, 2015

8.6K
In Vitro and In Vivo Detection of Mitophagy in Human Cells, C. Elegans, and Mice
08:40

In Vitro and In Vivo Detection of Mitophagy in Human Cells, C. Elegans, and Mice

Published on: November 22, 2017

17.6K

Area of Science:

  • Cell Biology
  • Autophagy Research
  • Organelle Quality Control

Background:

  • Mammalian cells possess mechanisms for peroxisome turnover, including Stub1-mediated pexophagy.
  • This pathway allows for cellular regulation of peroxisome number and integrity.
  • Heat shock protein 70 and the E3 ligase Stub1 are key components initiating pexophagy upon translocation to peroxisomes.

Purpose of the Study:

  • To detail methods for initiating and monitoring Stub1-mediated pexophagy using dye-assisted reactive oxygen species (ROS) generation.
  • To provide protocols for utilizing fluorescent proteins and synthetic fluorophores to trigger pexophagy in mammalian cell cultures.
  • To demonstrate the capability of targeting peroxisomes globally within a cell population or individually within single cells.

Main Methods:

  • Utilizing Stub1 (stress-induced protein 1) as an E3 ubiquitin ligase to mediate pexophagy.
  • Employing dye-assisted generation of reactive oxygen species (ROS) to activate the pexophagy pathway.
  • Implementing live-cell microscopy techniques to observe Stub1-mediated pexophagy dynamics.

Main Results:

  • Stub1 ligase activity facilitates the accumulation of ubiquitin and autophagy-related factors on targeted peroxisomes.
  • Elevated ROS levels within peroxisomes activate Stub1-mediated pexophagy.
  • Protocols using fluorescent dyes enable controlled initiation and monitoring of pexophagy in cell cultures.

Conclusions:

  • Stub1-mediated pexophagy is a viable pathway for controlling peroxisome homeostasis in mammalian cells.
  • Dye-assisted ROS generation offers a versatile tool to initiate and study pexophagy at both cellular and subcellular levels.
  • Live-cell imaging combined with dye-based ROS induction provides a powerful approach for investigating organelle turnover mechanisms.