| Gene name: | MIR185 |
| Aging type: | Accelerate |
| Aging characteristic: |
| Tissue type: | -- |
| Cell name: | HTC75,HFF,MRC5 |
| Experiment: | SA-β-gal activity assay//Immunofluorescence//TRF assay//qRT-PCR |
| Description: | Immunofluorescence and fluorescence in situ hybridization (IF-FISH) were performed in POT1 knockdown, miR-185 overexpression and POT1 rescue HTC75 cells with the indicatedγ-H2AX antibody and the TTAGGG telomere probe. Telomere restriction fragment (TRF) analysis showed the telomere length of HTC75 cells stably infected with the indicated viruses at the indicated population doublings. Either POT1 knockdown or miR-185 overexpression in HFF cells significantly increased CDKN1A (P21) and CDKN2A (P16) expression levels. Senescence-associated β-galactosidase (SA-β-gal) staining was performed to detect senescent cells. |
| Target gene: | POT1 |
| Official symbol(s): | POT1 |
| R-AG-Target gene: | Upregulation |
| Subcategory: | Unclear |
| Target gene experiment: | Dual-luciferase reporter assay//qRT-PCR//Western blot |
| Target gene description: | By using dual luciferase reporter assay, we compared the effects of miR-185 overexpression on wild-type POT1 (wPOT13’-UTR) or mutant POT1 (mPOT1-3’-UTR) luciferase activity. We stably transfected miR-185 into HTC75 cells and observed a decrease in POT1 mRNA levels. Consistent with the change in mRNA levels, POT1 protein levels were also decreased after miR-185 overexpression. |
| Regulatory pathway: | ATR |
| R-AG-Pathway: | Downregulation |
| Pathway experiment: | Western blot |
| Pathway description: | The total ATR protein level reduced upon miR-185 overexpression. |
Annotation:
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