| Gene name: | BTG3 |
| Aging type: | Prevent |
| Aging characteristic: |
| Tissue type: | -- |
| Cell name: | HCT116 |
| Experiment: | SA-β-gal activity assay//Flow cytometry//BrdU assay//Western blot//Cell morphological analysis//Knockdown |
| Description: | In contrast to what was observed previously in HCT116 cells,loss of BTG3 expression in IMR90 cells resulted in blunted cell proliferation, as demonstrated by a significant decrease of cells in the S phase, a moderate increase in the G1 population, a marked reduction in BrdU incorporation and RB phosphorylation. The slowed proliferation remained for at least for 2 weeks , indicating that this is not a transient growth arrest. Additionally, BTG3-knockdown cells displayed the enlarged, flattened morphology that is typical of senescent cells . The phenotype was confirmed by staining the cells for senescenceassociated β-galactosidase (SA-β-Gal) activity: we observed an increase in SA-β-Gal staining of at least sixfold 48 h after transfection with the BTG3-2 siRNA , which was further enhanced 1 week after transfection. |
| Target gene: | JMJD3 |
| Official symbol(s): | KDM6B |
| R-AG-Target gene: | -- |
| Subcategory: | Unclear |
| Target gene experiment: | RT-PCR//Knockdown |
| Target gene description: | Using RT PCR, we found that the expression of JMJD3 was increased in BTG3-knockdown cells. |
| Regulatory pathway: | ERK-AP1 |
| R-AG-Pathway: | -- |
| Official symbol(s): | MAPK1-JUN |
| Pathway experiment: | RT-PCR//SA-β-gal activity assay//Knockdown |
| Pathway description: | To confirm the involvement of the ERK signaling pathway, we treated BTG3-knockdown IMR90 cells with the ERK-specific inhibitor PD98059. As a result, a significant reduction in senescence was observed when ERK was inhibited. In contrast, two other unrelated drugs, SB202190 (p38 inhibitor) and SB202474 (negative control), had no apparent effect, indicating the specificity of the treatment. |
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