External factors
| External factors: |
senescence-messaging secretome factors |
| Experiment: |
Flow cytometry//SA-β-gal activity assay |
| Description: |
Young cells incubated in CM-old for a long time gradually acquired a senescence phenotype, including a flat morphology and cell hypertrophy. Using FACS analysis, we detected the marked increase in cell size after the continuous CM-old treatment for 7 days. The CM-old evoked a significant increase in the number of SA-β-Gal positive stained cells that is typical of senescent cells. |
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Regulatory relationship
| Regulatory pathway:
|
p16-MAPKAPK2-Rb//p53-p21-Rb |
| R-EF-Pathway:
|
Activation//Activation |
| Official symbol(s): |
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CDKN2A-MAPKAPK2-RB1//TP53-CDKN1A
|
| Pathway experiment: |
Western blot |
| Pathway description: |
The prolonged exposure of MESCs to CM-old resulted in long-term activation of H2A.X and ATM.Additionally, the immunoblot data were confirmed by immunofluorescence microscopy analysis for gH2A.X, pATM and p53BP1 localization. The phosphorylation levels of MAPKAPK-2 (a direct target of p38MAPK) and to a lesser degree p53 are increased in CM-treated cells with senescence progression. The senescent cells also displayed the elevated expression levels of both p21Cip1 and p16Ink4a whereas Rb phosphorylation was diminished gradually over all period of observation . |
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.