| Gene name: | BAZ1A |
| Aging type: | Prevent |
| Aging characteristic: |
| Tissue type: | -- |
| Cell name: | A549,U2OS |
| Experiment: | CCK-8 assay//Flow cytometry//SA-β-gal activity assay//Knockdown//EdU assay |
| Description: | CCK-8 assay showed BAZ1A-KD cells had decreased proliferation rate compared to control cells.In addition,cell cycle was arrested at G1 phase in BAZ1A-KD cells. Noteworthy, all of the five BAZ1A-KD cell lines showed increased percentage of positive SA-β-Gal stained cells. Moreover, reduced EdU incorporation rate was also observed in BAZ1A-KD cells , reflecting decreased level of DNA synthesis, which is also a well-known molecular phenotype of senescent cells. |
| Target gene: | SMARCA5 |
| Official symbol(s): | SMARCA5 |
| R-AG-Target gene: | -- |
| Subcategory: | Unclear |
| Target gene experiment: | Western blot//CCK-8 assay//Flow cytometry//SA-β-gal activity assay//Knockdown |
| Target gene description: | To examine whether SMARCA5 can regulate the abundance of BAZ1A and then influence senescence, we stably knocked down SMARCA5 by lentivirus mediated short hairpin RNA in A549 and U2OS cells, and reduced BAZ1A protein levels were found in SMARCA5-KD cells, which also exhibited senescenceassociated phenotypes, including decreased proliferation rate, cell cycle arrest at G1 phase, and increased percentage of positive SA-β-Gal stained cells. |
| Regulatory pathway: | SMAD3-P21 |
| R-AG-Pathway: | -- |
| Official symbol(s): | SMAD3-CDKN1A |
| Pathway experiment: | ChIP-qPCR//qRT-PCR//Western blot |
| Pathway description: | The results showed enriched signal of BAZ1A binding to the promoter region of SMAD3 compared to non-specific IgG binding control, which was further validated by Chromatin Immunoprecipitation coupled with quantitative PCR (ChIP-qPCR) .Our data showed the upregulation of both SMAD3 and CDKN1A in BAZ1A-KD cells,indicating upregulated SMAD3 activated the expression of CDKN1A and led to slower cell proliferation and ultimately cellular senescence. |
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