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A Quantitative Measurement of Reactive Oxygen Species and Senescence-associated Secretory Phenotype in Normal Human Fibroblasts During Oncogene-induced Senescence
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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
Intracellular ROS has been shown to play an important role in the induction of cellular senescence. Here, we describe a sensitive assay for quantifying ROS levels during cellular senescence. We also provide protocols for assessing the senescence-associated secretory phenotype, which reportedly contributes to various age-related dysfunctions.
