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Figure 5 | Molecular Cancer

Figure 5

From: PTEN suppresses the oncogenic function of AIB1 through decreasing its protein stability via mechanism involving Fbw7 alpha

Figure 5

Effect of PTEN on the transcriptional activity of AIB1. (A) COS-7 cells were transfected with Gal4-luc reporter construct and Gal4-DBD-tagged AIB1 or VP16 together with wt or mu PTEN. pcDNA3.1(−)/pcGal4-DBD vector was used as empty vector (EV). Twenty four hours after transfection, the cells were collected and subjected to luciferase activity assay. (B) COS-7 cells were transfected with Gal4-luc reporter construct and Gal4-DBD-tagged AIB1 together with increasing dosages (10, 100, 300 ng) of Gfp-tagged wt or mu PTEN. pcDNA3.1(−)/pcGal4-DBD vector was used as EV. Twenty four hours after transfection, the cells were collected and subjected to luciferase activity assay. (C) COS-7 cells were transfected with Gal4-luc reporter construct and Gal4-DBD-tagged AIB1 together with or without Gfp-tagged PTEN. pcDNA3.1(−)/pcGal4-DBD vector was used as EV. The cells were treated with or without 10 μM LY294002 for 12 h, and then subjected to luciferase activity assay. (D & E) COS-7 cells were transfected with Gal4-luc reporter construct and Gal4-DBD-tagged AIB1 together with various combinations of other constructs as indicated. pcDNA3.1(−)/pcGal4-DBD vector was used as EV. Twenty four hours after transfection, the cells were harvested and subjected to luciferase activity assay. (F) MCF-7 cells were transfected with ERE-luc reporter and ERα constructs along with other plasmids as indicated. pcDNA3.1(−) vector was used as EV. Eight hours after transfection, the cells were switched to phenol red-free medium containing 10% charcoal-dextran-treated fetal bovine serum for 16 h followed by treatment with or without 10 nM E2 for another 16 h, and then subjected to luciferase activity assay. (G) MCF-7 cells were transfected with Gfp-tagged wt or mu PTEN. Eight hours after transfection, the cells were switched to phenol red-free medium containing 10% charcoal-dextran-treated fetal bovine serum for 16 h followed by treatment with or without 10 nM E2 for another 16 h. The cells were then subjected to real-time PCR to measure the mRNA level of pS2 and c-myc. Each bar represents the mean ± S.D. from three independent experiments. *, P < 0.05; **, P < 0.01.

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