| Gene name: | ACLY |
| Aging type: | Prevent |
| Aging characteristic: |
| Tissue type: | -- |
| Cell name: | HDF |
| Experiment: | Knockdown//Cell morphological analysis//Cell proliferation assay//SA-β-gal activity assay |
| Description: | ACLY knockdown cells showed apparent growth retardation and a typical senescent morphology (flat and enlarged in size). The proliferation assay and senescence-associated -β-galacto-sidase staining showed that knockdown of ACLY in HDF cells significantly reduced their proliferation rate. A large proportion of ACLY knockdown cells were positive for senescence-associated-β-galactosi-dase staining. |
| Regulatory pathway: | P53-AMPK |
| R-AG-Pathway: | -- |
| Official symbol(s): | TP53-PRKAA1 |
| Pathway experiment: | Knockdown//SA-β-gal activity assay//qRT-PCR//DAPI staining//Immunofluorescence//Pull-down assay |
| Pathway description: | Initially, we silenced p53 in ACLY knockdown HDF cells, and found that additional knockdown of p53 completely abrogated ACLY silencing-induced cellular senescence. we observed an increased p53 protein level in ACLY knockdown HDF cells by confocalmicroscopy and increased mRNA levels of p53 downstream target genes by real-time quantitative PCR.The direct interaction of ACLY with AMPK a2 was confirmed by an in vitro glutathione S-transferase(GST) pulldown assay . Immunofluorescent staining of over-expressed AMPK and endogeneous ACLY showed that about 26% and 37% of the signal co-localized in HDF and HCT116 cells, respectively .An in vitro kinase assay using immunoprecipitated AMPK a2 showed an apparent reduction of AMPK activity by ACLY. |
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