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. 2013 Apr 11;4(4):e583. doi: 10.1038/cddis.2013.98

Figure 4.

Figure 4

Epigenetic regulation of IGFBP2 is mediated by menin via HDAC. (a) Menin represses IGFBP2 promoter activity. The 293T cells were transfected with the IGFBP2 promoter-luciferase reporter and epitope-tagged menin or empty vector control. After incubation for 24 h, luciferase activity was measured. Each transfection was performed in duplicate and repeated three times. The means±S.D. of duplicate determinations from three separate experiments are shown. The expression level of transfected menin is shown at the bottom. (b) Increased level of genomic IGFBP2 expression in Men1−/− MEFs was confirmed using qRT-PCR. (c) Men1+/+ MEFs cells were treated with siRNA targeting SUV39H1. The expression level of IGFBP2 was analyzed by qRT-PCR. Error bars represent S.D., n=3. Significance of differences was evaluated (*P-value<0.05). (d and e) Men1+/+ and Men1−/− MEF cells were treated with TSA for 24 h with increasing dose as indicated. Total RNA was prepared to perform qRT-PCR. The expression level of IGFBP2 and GBX2 were normalized by GAPDH. TSA-dependency was shown as the fold difference between samples by considering expression levels obtained from each Men1+/+ and Men1−/− MEF cells without TSA treatment as 1, respectively. Error bars represent S.D., n=3 (*P-value<0.05)