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Loss of Proteostasis

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Hallmarks of Aging Miniseries No. 4 Loss of Proteostasis

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As cells age, environmental stresses add up and mechanisms responsible for maintaining proper protein composition start to decline. Proteins lose their stability, autophagic processes start to fail, and misfolded proteins accumulate.

Over the years, our bodies are subjected to many environmental inputs that put thermal stress, oxidative stress, and osmotic stress on our cells, causing misfolding of proteins. For example, free radicals present in polluted air have been identified as particularly noxious agents in this regard, contributing to multiple aging-related pathologies. In younger cells, micro- and macroautophagy pathways, together with the ubiquitin-proteasome system, take care of clearing these unfolded proteins. However, in aging cells, autophagy induction can be gradually compromised, and lysosomes become less efficient at eliminating the vesicles carrying this cellular waste.

In a vicious cycle termed “inflammaging,” this decrease in efficient autophagy results in an increase in intracellular ROS, triggering the damage-sensing inflammasome to generate low levels of chronic inflammation, further accelerating aging.

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Featured Solution for Studying Loss of Proteostasis
The GFP-LC3 displays a diffuse nuclear and cytosolic distribution in fed cells, and a punctate distribution in starved autophagic cells.
HeLa cells were transduced with LentiBrite GFP-LC3 lentiviral particles and were either (A) left in complete media or (B) incubated in EBSS containing a lysosome inhibitor under starvation conditions. The GFP-LC3 displays a diffuse nuclear and cytosolic distribution in fed cells, and a punctate distribution in starved autophagic cells. (Click to enlarge)
LentiBrite™ Lentiviral Biosensors for LC3 and p62 with GFP and RFP MilliporeSigma’s LentiBrite™ Lentiviral Biosensors, a new suite of pre-packaged lentiviral particles encoding foundational proteins of autophagy detection - LC3 and p62, enabling precise visualization of autophagosome formation under different cell/disease states in live cell and in vitro analysis. Visualize autophagy in real time, even in difficult-to-transfect cell types, using LentiBrite™ GFP- & RFP-tagged LC3 and p62 wild-types and LC3-G120A mutant control lentiviral biosensors. 

Advantages:

  • Pre-packaged, ready-to-use, fluorescently-tagged LC3 & p62 with monomeric GFP & RFP
  • Minimum titer (≥3 x 10^8 IFU/mL) per vial
  • Long-term, stable fluorescent expression that is non-disruptive towards cellular function
  • Higher efficiency transfection as compared to traditional chemical-based and other non-viral-based transfection methods
  • Ability to transfect dividing, non-dividing, and difficult-to-transfect cell types, such as primary cells or stem cells
  • Validated for fluorescent microscopy and live cell analysis
  • LC3 Control Mutant lentiviral particle contains the translocation-defective protein LC3-G120A for comparison studies.

LentiBrite GFP-LC3 Lentiviral Biosensor (Catalogue No. 17-10193)

Timecourse imaging of HT-1080 cells after transduction with LentiBrite GFP-LC3 and the addition of a lysosomal inhibitor under starvation conditions. GFP-LC3 displays a diffuse nuclear and cytosolic distribution in fed cells, and a punctate distribution in starved, autophagic cells.



DescriptionCat. No.
LentiBrite™ GFP-LC3 Lentiviral Biosensor 17-10193
LentiBrite™ RFP-LC3 Lentiviral Biosensor 17-10143
LentiBrite™ GFP-LC3 Control Mutant Lentiviral Biosensor 17-10189
LentiBrite™ RFP-LC3 Control Mutant Lentiviral Biosensor 17-10188
LentiBrite™ GFP-p62 Lentiviral Biosensor 17-10224
LentiBrite™ RFP-p62 Lentiviral Biosensor 17-10404


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