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. 2017 Mar 12;8(5):894–902. doi: 10.7150/jca.17064

Figure 4.

Figure 4

TET1 mediates GPER up-regulation by hydroxymethylation in insulin-driven microenvironment. A. Ishikawa and HEC-1A cells were treated with 100nM insulin for 24 h. GPER transcriptional level was analyzed by real time PCR. Data shown were normalized for GAPDH expression and then calculated as fold changes of relative mRNA expression. B. Insulin induced GPER expression through TET1. Endometrial cancer cells were transfected by siCON or siTET1. Cells with or without silencing TET1 were treated with 100nM insulin for 48h. The fold change of GPER relative to control group was calculated from densitometry. C. Increased TET1 expression up-regulates GPER expression. TET1 was overexpressed in endometrial cancer cells by tranfecting pPB-TET1 plasmid. GPER and TET1 expression were analyzed by western blot. D. TET1 affects hydroxymethylation of GPER gene promoter in insulin condition. HEC-1-A cells transfected with siTET1 or siCon were treated with 100nM insulin for 24h. 5-hmC level of GPER gene promoter fragment was detected by hMeDIP. * p < 0.05, **p < 0.01,***p < 0.001 compared to control.