| Gene name: | HDAC4 |
| Aging type: | Prevent |
| Aging characteristic: |
| Tissue type: | -- |
| Cell name: | 2BS |
| Experiment: | Knockdown//Western blot//qRT-PCR//MTT assay//Flow cytometry//SA-β-gal activity assay |
| Description: | The overexpression and knockdown of HDAC4 were confirmed by western blot. Overexpression of HDAC4 resulted in only scattered senescence-associated β-galactosidase (SA-β-gal) activity, which is a biomarker for senescent cells. The formation of senescence-associated heterochromatin foci (SAHF), which is another hallmark of senescent cells, was not observed when HDAC4 was overexpressed. Continuous cell growth and a decrease in the proportion of 2BS cells in the G0/G1 phases were evident in HDAC4 overexpressing cells compared with corresponding empty control vector-infected cells. In contrast, HDAC4 knockdown induced more robust senescence phenotypes, which displayed an elevated SA-β-gal activity, prominent heterochromatin foci indistinguishable from those in senescent 2BS cells, pronounced cell growth arrest, and accelerated G1 cell cycle arrest compared with corresponding empty control vector-infected cells. |
| Target gene: | SIRT1 |
| Official symbol(s): | SIRT1 |
| R-AG-Target gene: | -- |
| Subcategory: | Unclear |
| Target gene experiment: | Knockdown//RT-PCR//Western blot//IP |
| Target gene description: | We first analysed the expression patterns of SIRT1 using western blot and RT-PCR subjected to such perturbations.The protein levels of SIRT1 were markedly enhanced in HDAC4-transfected HeLa cells compared with the vector-transfected cells. In addition, we silenced HDAC4 expression in HeLa cells. Not unexpectedly, there was a marked decrease in the SIRT1 protein level in these HDAC4 silenced cells.Moreover,the RT-PCR analysis demonstrated no alteration at the SIRT1 mRNA levels in either HDAC4-overexpressing or knockdown cells, indicating that HDAC4 might normally function to increase the SIRT1 protein levels, but not at the mRNA level.To gain further insight into the interplay between HDAC4 and SIRT1, cellular interactions between HDAC4 and SIRT1 were confirmed using immunoprecipitation (IP) assays. |
| Regulatory pathway: | -- |
| R-AG-Pathway: | -- |
| Pathway experiment: | -- |
| Pathway description: | -- |
Annotation:
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