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En kvantitativ måling af reaktive ilt arter og gulne-associerede sekretoriske fænotype i normale humane fibroblaster under onkogen-induceret gulne
JoVE Journal
Developmental Biology
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JoVE Journal Developmental Biology
A Quantitative Measurement of Reactive Oxygen Species and Senescence-associated Secretory Phenotype in Normal Human Fibroblasts During Oncogene-induced Senescence
DOI:

13:59 min

August 12, 2018

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Chapters

  • 00:00Title
  • 01:10Inducing Oncogene-induced Senescence
  • 03:11Monitoring Senescence by Senescence Associated ß-galactosidase (SA ß-gal) Staining
  • 04:27Quantifying ROS Induction during OIS
  • 06:42Quantifying IL-6 and IL-8 mRNA Expression for SASP Anaylsis using Real-time PCR
  • 08:53Quantifying the Levels of Secreted IL-6 and IL-8 Proteins for SASP Analysis using ELISA
  • 10:56Results: Representative ROS and SASP Expression Analysis during H-Ras-induced Senescence
  • 12:25Conclusion

Summary

Automatic Translation

Intracellulære ROS har vist sig at spille en vigtig rolle i induktion af cellulære ældning. Her, beskriver vi en følsom analyse for kvantificering ROS niveauer under cellulære ældning. Vi leverer også protokoller for at vurdere den gulne-associerede sekretoriske fænotype, som efter sigende bidrager til forskellige aldersrelaterede dysfunktioner.

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