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. 2013 Dec 20;8(12):e83312. doi: 10.1371/journal.pone.0083312

Figure 3. CHIP regulated FoxO3 binding to bim and overexpression of CHIP induced apoptosis resistance.

Figure 3

(A) CHIP inhibited FoxO3 binding to the bim promoter. MCF7 and MCF10A cells were transfected with or without Myc-CHIP, followed by ChIP assay. The relative band intensity of PCR products revealed the binding of FoxO3 to the bim promoter. RT-PCR was performed for bim promoter (* P<0.05). (B) Knockdown of CHIP increased bim transcription in both MCF7 and MCF10A cells. Cells were transfected with CHIP siRNA or negative control siRNA, followed by ChIP assay. The relative band intensity of PCR products revealed the binding of FoxO3 to the bim promoter. RT-PCR was performed for bim promoter (* P<0.05). (C) Overexpression of CHIP attenuated apoptosis in cisplatin-treated MCF7 and MCF10A cells. Myc-CHIP was overexpressed in MCF7 and MCF10A cells that were subsequently treated with cisplatin for 24 h. All groups of cells were analyzed by flow cytometry (* P<0.05).