External factors
| External factors: |
Hypoxia |
| Experiment: |
SA-β-gal activity assay//Immunofluorescence//BrdU assay//SAHF//Western blot |
| Description: |
Indeed, compared to normoxia (20% O2) in hypoxia we observed reversal of H-RasV12- driven senescence induction as shown by negative staining of the cells for SA-β-gal activity.We found that HDFs ectopically expressing H-RasV12 were positive for Ki67 antigen and incorporated BrdU to a higher extent under low oxygen conditions when compared to normoxia.We also tested whether H-RasV12 overexpression results in generation of SAHFs, and here we showed that SAHF formation takes place only under normoxic conditions but not when the cells were cultured under hypoxic conditions.We found that the cells grown under hypoxic conditions have reduced protein levels of all of the senescence hallmarks tested including p53, p16INK4a, p21CIP1 and HP1γ. |
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Regulatory relationship
| R-EF-Target gene:
|
Activation//Upregulation |
| Official symbol(s): |
HIF-1A//MIF |
| Target gene experiment: |
Western blot//RT-PCR |
| Target gene description: |
As shown by protein analyses as well as mRNA expression levels, indeed, stabilization of HIF-1a was detected in both cell lines in hypoxia but not in normoxia .Thus we also assessed MIF expression in the same setting and detected a modest increase in MIF protein as well as mRNA levels under the hypoxic conditions. |
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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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