| Gene name: | MYC |
| Aging type: | Accelerate |
| Aging characteristic: |
| Tissue type: | -- |
| Cell name: | RPE |
| Experiment: | SA-β-gal activity assay//Knockdown |
| Description: | The SA-β-gal activity assay was performed on these two cell clones with or without 4-OHT treatment on Day 24. With the addition of 4-OHT, the average number of SA-β-gal-positive cells per unit area was increased in both the cells with LL-c-Myc (clone 4) and ML-c-Myc (clone 3). In order to determine whether long-term post-confluent cells entered into senescence rather than cellular quiescence, we replated the confluent RPE LL-c-Myc cells (clone 4) treated with OHT for 24?days, and measured their SA-β-gal activity at different time points. The percentage of β-gal-positive cells after 2 and 6?days was 85.6 and 87.3%, respectively, suggesting that the replated cells were not proliferating, and the confluent cells were substantially senescent rather than quiescent. Collectively, we concluded that low level of c-Myc expression (LL-c-Myc) induced cellular senescence rather than quiescence in post-confluent RPE cells during long-term culture.After infecting with lentivirus expressing shRNA, all the cells expressing high level of c-Myc (HL-c-Myc) eventually died after 8?days, while HL-c-Myc cells infected with scrambled lentiviral shRNA control could survive for 12?days. For ML-c-Myc cells,it took 24?days for the cells infected with scrambled lentiviral shRNA to die completely. |
| Target gene: | AP4//P53 |
| Official symbol(s): | REPIN1//P53 |
| R-AG-Target gene: | --//-- |
| Subcategory: | Unclear |
| Target gene experiment: | Knockdown//Western blot |
| Target gene description: | The expression of AP4 increased evidently in the samples that incubated with 4-OHT for the initial 14?h, while c-Myc-ER level was relatively high. Both AP4 and c-Myc-ER protein were subsequently decreased during longer incubation, which might probably due to protein degradation. After normalized to the signal of actin, the expression of AP4 increased sevenfold during the first 14?h upon the activation of c-Myc,suggesting that RPE c-Myc-ER clones were functional and could be used for further studies.Western blot analysis showed that differential c-Myc expression in the cell clones induced a corresponding level of AP4 protein.In the cells with LL-c-Myc, the expression of p53 decreased more with a larger decrease in AP4 level, indicating that p53 expression was regulated by AP4, which is consistent with our previously finding .In this study, knockdown of AP4 expression brings about a decrease in p53 expression, but the decrease in the expression of p53 was weak in RPE cells when c-Myc level is higher. |
| Regulatory pathway: | -- |
| R-AG-Pathway: | -- |
| Pathway experiment: | -- |
| Pathway description: | -- |
Annotation:
Loading,please wait...