| External factors: | BrdU |
| Aging type: | Accelerate |
| Aging characteristic: |
| Category: | Chemical compounds |
| Phenotype: | Aging |
| Experiment: | SA-β-gal activity assay//Cell morphological analysis |
| Description: | Senescence associated morphological changes were detected within 48 hours and treated cells show flatter, enlarged and granule rich morphology,similar to morphology of replicatively senescent cells reported previously. The senescent state of BrdU treated cells was also confirmed using SA-β-gal, the most widely used assay for cellular senescence. |
| Regulatory pathway: | DDR-ROS-ATM |
| R-EF-Pathway: | -- |
| Official symbol(s): | DDR-ROS-ATM |
| Pathway experiment: | Western blot//53BP1 foci formation assay |
| Pathway description: | Robust proliferation arrest and activation of ATM kinase was detected in cells exposed to BrdU after 48 hours, monitored by the presence of phosphorylation of H2Ax protein, a substrate of ATM kinase. Towards this, senescent cells generated by BrdU exposure were treated with ATM kinase inhibitor caffeine or Ku53393 or ROS quencher NAC after 48 hours as described in previous section. As an indicator of ATM kinase inhibition, the presence of 53BP foci was evaluated in these cells, which was found to be significantly lowered when ATM kinase was inhibited , but not in ROS quenched cells. |
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