External factors
| External factors: |
Bleomycin |
| Category: |
Chemical compounds |
| Experiment: |
SA-β-gal activity assay//RT-PCR//Western blot |
| Description: |
We exposed presenescent human fibroblasts (strain HCA2) to a senescenceinducing dose of bleomycin, which generates extensive DNA damage. RT-PCR showed that, relative to untreated presenescent controls, transcripts encoding both proteins increased steadily over 7 days. Quantitative real time PCR (qPCR) confirmed a 6- to 7-fold senescence-associated increase in these transcript levels. The gradual increase over several days matched the kinetics with which the SASP develops (10). In these and subsequent experiments, we confirmed senescence by growth arrest (not shown), morphology, and staining for senescence-associated -galactosidase (SA-β-gal),Both proteins were barely detectable in presenescent cells, but the levels increased significantly when cells became senescent. |
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Aging network
Annotation:
The green line represents Upregulation.
The purple line represents Downregulation.
The orange line represents Activation.
The yellow line represents Inhibition.
The gray line represents Unclear.
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