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

Figure 4. CHIP modulated PTEN, and knockdown of PTEN inhibited the AKT/FoxO3a/Bim signaling pathway.

Figure 4

(A) CHIP overexpression reduced the synthesis of PTEN mRNA, and knockdown of CHIP increased PTEN mRNA level. MCF7 and MCF10A cells were transfected with Myc-CHIP, Myc empty vector, CHIP siRNA or negative control siRNA respectively. Total cellular mRNA was then extracted and subjected to reverse transcription PCR. The PTEN mRNA level was determined and calculated from three independent experiments (P<0.05). (B) CHIP regulated PTEN protein level in both transcriptional and post-translational steps. MCF7 and MCF10A cells were transfected with or without Myc-CHIP. After 48 h, the cells were treated for western blot, ChIP assay (* P<0.05) or in vivo ubiquitylation assay respectively. (C) Knockdown of PTEN promoted AKT activation and influenced the AKT/FoxO3/Bim signaling pathway. MCF7 and MCF10A cells were transfected with PTEN siRNA or negative control siRNA. The altered expression patterns of p-AKT (T308), AKT, p-FoxO3, FoxO3, Bim, cleaved caspase 9, and cleaved PARP were determined by western blot.