Gene name: | MAD2L1 |
Aging type: | Prevent |
Aging characteristic: |
Tissue type: | -- |
Cell name: | IMR-90,MCF 10A |
Experiment: | SA-β-gal activity assay//Immunostaining//SAHF |
Description: | Strikingly, several IMR90- siMAD2 cells were (SA)-β gal-positive (pH 6, 58%). The percentage of positive cells forgH2AX was higher (about 60%) in MCF10A-siMAD2 cells than in MCF10A-siGFP control cells .We then looked for SAHFs presence in primary human fibroblasts after 30 days from siMAD2 transfection and we scored that almost all of the cells (about 85%) showed heterochromatin foci when stained with DAPI. |
Target gene: | P14 |
Official symbol(s): | P14 |
R-AG-Target gene: | -- |
Subcategory: | Unclear |
Target gene experiment: | Western blot//qRT-PCR |
Target gene description: | At 72 h post-transfection, p14ARF mRNA was highly increased in IMR90 human fibroblasts. Similarly, Western-blotting experiments showed a high increase of p14ARF protein levels in IMR90- siMAD2 cells, suggesting that p14ARF could mediate the senescence response in these cells by sensing aneuploidy. |
Regulatory pathway: | P53-P21 |
R-AG-Pathway: | -- |
Official symbol(s): | TP53-CDKN1A |
Pathway experiment: | Western blot//qRT-PCR |
Pathway description: | Real-time RT-PCR revealed a high increase (sevenfold) of p21waf1 mRNA in primary human fibroblasts with MAD2 haploinsufficiency when compared to IMR90-siGFP control cells . As expected Western blotting confirmed that p21waf1 protein was significantly higher in siMAD2 than in IMR90-siGFP transfected cells. However, by Western blot analysis we detected elevated levels of p53 protein suggesting that the observed p21waf1 accumulation could be induced by p53 trans-activation of the p21waf1 gene. When p53 was posttranscriptionally silenced in IMR90-siMAD2 cells, expression level of the p21waf1 gene resulted similar to that present in IMR90-siGFP control cells, as assessed by real-time RT-PCR analysis. |
Annotation:
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