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. 2015 Oct 8;113(8):1186–1196. doi: 10.1038/bjc.2015.273

Figure 6.

Figure 6

Effects of FOXO1 silencing on HER2 expression in GC cells and xenograft tumours. (A) FOXO1 was downregulated by transfection of FOXO1 shRNA (shFOXO1) into SNU-638 and MKN45 cells. Immunoblot analysis (WB) and RT–PCR showed that FOXO1 shRNA transfection increased the expressions of HER2 protein and mRNA compared with control shRNA (shCtrl) transfection. (B) FOXO1 expression in SNU-216 cells was upregulated by transfection of FOXO1A3. WB and RT–PCR showed that FOXO1A3 transfection decreased HER2 protein and mRNA expressions compared with pcDNA3 transfection. (C) ChIP for the binding of FOXO1 to the HER2 promoter in SNU-638 cells. Chromatin was cross-linked and immunoprecipitated using anti-FOXO1. Precipitated DNAs were amplified and quantified by real-time PCR using two primers (PM1 and PM2). The results are expressed as percentages of the input level. Bars represent mean±s.d. (n=3). *P<0.05, compared with shCtrl. (D) Representative features of immunohistochemical staining for FOXO1 (upper panels) and HER2 (lower panels). Sections obtained from orthotopic xenograft tumours showed that shFOXO1-expressing tumours had higher HER2 expression than shCtrl-expressing tumours ( × 400 magnification).