External factors
| Category: |
Chemical compounds |
| Cell name: |
HT-1080,B16F10 |
| Experiment: |
SA-β-gal activity assay//Western blot//Cell morphological analysis |
| Description: |
While assaying the antiproliferative effects of E235 on transformed cells, we observed that cells treated for at least 48h with lower doses of E235 (0.25-1μM) increased in size and displayed characteristics consistent with a senescent phenotype. Both 15 HT1080 and B16F10 cells exhibited a dramatic increase in perinuclear SA-β-gal staining after 48h of treatment with lower doses of E235, with almost all of the cells treated with 1μM E235 being positive for β-gal expression.Both HT1080 and B16F10 cells displayed elevated levels of p21 with 1μM E235 at 8 and 16h. |
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Regulatory relationship
| R-EF-Target gene:
|
Upregulation |
| Target gene experiment: |
Luciferase reporter assay//Western blot |
| Target gene description: |
E235 caused a dose-dependent increase in luciferase activity from a reporter plasmid containing the 5 UTR of the ATF4 mRNA fused to the luciferase gene .A dose-dependent increase in ATF4 protein levels was seen only in the HT1080 and B16F10 cells. There was no appreciable induction of ATF4 protein in the normal diploid AG1522 human fibroblasts in response to E235, whereas thapsigargin induced a potent upregulation of ATF4 levels , suggesting that this effect is restricted to transformed cells. |
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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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