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. 2016 Mar 24;6:23665. doi: 10.1038/srep23665

Figure 5. Metformin regulates PPARα-mediated Igfbp-2 gene expression.

Figure 5

(A) Effect of the Pparα on mouse Igfbp-2 promoter activity. Transient transfections were performed using observed promoter construct, Igfbp-2 (−1314/+78) in 293T cell. Below, immunoblot analysis of Sirt1 and Pparα expression in transfected HEK293T cells, detected with anti-Sirt1 and PPARα. Equal amounts of protein were confirmed using β-actin. (B) Schematic drawing of Pparα binding region of Igfbp-2 promoter. Serial deletion constructs of Igfbp-2 promoter (−698/−686 bp) were transiently transfected with Pparα to 293T cell. (C) PPRE-dependent activation of Igfbp-2 promoter activity. Alignment of potential PPRE in Igfbp-2 promoter is indicated (top panel). (D) ChIP assay. Mouse primary hepatocytes were treated metformin for 24 h. Chromatin was precipitated using an anti-PPARα antibody from primary hepatocytes, and purified DNA samples were used to perform qPCR with primers binding to the PPRE regions on the Igfbp-2 gene promoter. Input represents 10% of purified DNA in each sample. *p < 0.05 vs. untreated control or individual-treated cells. mIgfbp-2, mouse Igfbp-2.