| Gene name: | MCRS1 |
| Aging type: | Accelerate |
| Aging characteristic: |
| Tissue type: | -- |
| Cell name: | IMR-90,Hs68 |
| Experiment: | PI staining//SA-β-gal activity assay//Cell morphological analysis |
| Description: | To confirm that MSP58-overexpressing cells entered senescence, we analyzed DNA content using flow cytometry with propidium iodide staining. HT1080 MSP58 clone 11 and 14 cells showed decreased fractions in the G1phase resulting from cells accumulating in the S and G2/M phases and elevated forward and side scatter relative to control cells, respectively reflecting increased cell size and granularity.Notably, stable HT1080 and NIH3T3 transfectants expressing MSP58 showed elevated SA-β-gal activity, a classical biomarker of senescence.After selection, MSP58-infected cells displayed a large size with a flattened shape and significantly increased SA-β-gal activity compared with control cells. |
| Target gene: | BRG1 |
| Official symbol(s): | SMARCA4 |
| R-AG-Target gene: | -- |
| Subcategory: | Unclear |
| Target gene experiment: | -- |
| Target gene description: | -- |
| Regulatory pathway: | P53-P21 |
| R-AG-Pathway: | Activation |
| Official symbol(s): | TP53-CDKN1A |
| Pathway experiment: | Luciferase reporter assay//Western blot//SA-β-gal activity assay |
| Pathway description: | Elevated levels of p53 and p21 proteins were also detected in MSP58-overexpressing IMR90, Hs68, and H184B5F5/M10 cells;Cells transfected with the truncated constructs showed substantially reduced responses as the p21-dm construct was not induced, indicating that MSP58 regulates p21 promoter activity through the p53 protein.Furthermore, depletion of ATM or p21 using shRNA transfection significantly reduced the incidence of MSP58-induced senescent phenotypes as shown by the representative stable clones HT1080 MSP58 clone 11-ATMsi 9 and MSP58 clone 11-p21si6. |
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