| Gene name: | KLF4 |
| Aging type: | Accelerate |
| Aging characteristic: |
| Tissue type: | -- |
| Cell name: | T-REx-293 KLF4,T-REx-HeLa KLF4 |
| Experiment: | SA-β-gal activity assay//Western blot |
| Description: | Then we characterized senescence in KLF4 overexpressed cells. Senescenceassociated β-galactosidase activity (SA-β-gal) markedly increased in cells with KLF4 overexpression . We also detected the senescence related proteins by Western Blot, and found that p21W AF1/CIP1 protein was strongly accumulated after KLF4 overexpression, accompanied by reduced expression of Cyclin E1 and increased expression of Suv39H1, but no significant variation of p53 and p16Ink4a. |
| Target gene: | P21 |
| Official symbol(s): | P21 |
| R-AG-Target gene: | Upregulation |
| Subcategory: | Unclear |
| Target gene experiment: | CHIP//qPCR//Western blot |
| Target gene description: | We found that p21 mRNA level was induced by KLF4 overexpression, and KLF4 could bind to the promoter region of p21 gene, confirmed by ChIP assay. When p21 protein was knocked down, KLF4 induction could induce only about 8 percent of senescent cells, comparing with more than 70% senescent cells in control cells . |
| Regulatory pathway: | MIR-203-SURVIVIN-P12 |
| R-AG-Pathway: | -- |
| Official symbol(s): | BIRC5-POLD4 |
| Pathway experiment: | Western blot//qPCR//Luciferase reporter assay |
| Pathway description: | Protein level of survivin and mRNA expression were both inhibited by KLF4 overexpression. Additionally, p21 upregulation induced by KLF4 was significantly inhibited.In our study, survivin protein could directly bind to the distal and proximal p53 binding sites of p21 promoter in T-REx-293 KLF4 cells, as confirmed by ChIP assay. T-REx-293 cells were co-transfected with survivin and reporter plasmids, and reporter assay showed that the transcription activities of both pGL3 p21 5′, pGL3 p21 3′ were significantly inhibited by survivin. ChIP assay showed that KLF4 could directly bind to -189bp to +11bp fragment of miR-203 gene. To test whether miR-203 can target survivin or not, pre-miR-203 precursor and miR-203 inhibitor were transfected into T-REx-293 cells, respectively.mRNA expression of survivin was inhibited by pre-miR-203, while inhibition of miR-203 could increase survivin expression, the similar result was shown in protein level. |
Annotation:
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