Effect of G9a on the transcription and protein expression of Sox2 in various cell types.

<p>(A) The effect of ectopic expression of G9a on Sox2 and Oct4 protein expression in three breast cancer cell lines (MCF7, ZR-75-1, and MDA-MB-231) and two glioblastoma cell lines (U87-MG and T98-G) was determined by immunoblot analysis. Each cell line was transfected with a G9a-expressing plasmid for 24 h and then harvested for analysis. For quantitative analysis of immunoblotting results, the mean density of the Sox2 band was measured with Multi Gauge V3.0 software, and the band density was divided by the density of β-actin to obtain the normalized band density. (B) Cells were transfected with control or G9a expressing plasmids for 24 h, and the effect of ectopic expression of G9a on the transcription of Sox2 was quantitatively analyzed by real-time RT-PCR. (C) The effect of G9a activity on Sox2 protein stability in mouse ESCs. Mouse ESCs were treated with 0.2 μM BIX-01294 for the indicated times and then harvested for immunoblot analysis. (D) The effect of G9a activity on the transcription of Sox2 in mouse ESCs. Mouse ESCs were treated with 0.2 μM BIX-01294 for the indicated times and then harvested for quantitative analysis of Sox2 transcription by real-time RT-PCR. Abbreviations: G9a, G9a-expressing plasmid; Sox2, Sox2-expressing plasmid; C, control plasmid. *P < 0.05 compared with the control group. Data are expressed as the mean ± SD and are representative of three independent experiments.