| Gene name: | ING1 |
| Aging type: | Accelerate |
| Aging characteristic: |
| Tissue type: | -- |
| Cell name: | Fibroblast |
| Experiment: | SA-β-gal activity assay |
| Description: | ING1a induced the expression of both p16 and Rb when ectopically expressed in young fibroblasts. ING1a also induced senescence-associated β-galactosidase staining and cell cycle arrest at the G0/G1phase of the cell cycle after about 48 h of ectopic expression. |
| Target gene: | ITSN2 |
| Official symbol(s): | ITSN2 |
| R-AG-Target gene: | Upregulation |
| Subcategory: | Unclear |
| Target gene experiment: | RT-PCR//BrdU assay//Knockdown//SA-β-gal activity assay |
| Target gene description: | We measured the expression of ITSN2 in senescent cells after knocking down ING1a using siRNA. We found that knockdown of ING1a in senescent cells led to significant down-regulation of ITSN2 mRNA levels, further suggesting a role for ING1a in regulating ITSN2 expression .When ITSN2 was knocked down in ING1a-expressing cells, we found that EGFR internalization kinetics were similar to control GFP-expressing cells and were significantly restored when compared to the ING1a- expressing A431 cells. We next checked if loss of ITSN2 in ING1a-expressing cells affected the senescence phenotype. We found that senescence-associated β-gal staining was reduced significantly in these cells, and they showed increased proliferation when assayed using the BrdU incorporation assay. High levels of p16 were no longer induced by ING1a , suggesting that ITSN2 was indeed a direct mediator of ING1a-induced senescence signal. Since we previously noted higher levels of both ING1a and ITSN2 in senescent versus low passage fibroblasts. |
| Regulatory pathway: | RB-E2F |
| R-AG-Pathway: | -- |
| Official symbol(s): | RB1-E2F1 |
| Pathway experiment: | Western blot |
| Pathway description: | Since hypophosphorylated Rb is the Activate form that binds and inhibits E2F, we next asked whether the Rb in ING1a-expressing cells physically associated with E2F. Western blot analysis of immunoprecipitated E2F1 in these samples confirmed that E2F bound Rb, avidly in the presence of ING1a compared to the GFP-expressing cells. |
Annotation:
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