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. 2014 Sep 25;5(21):10584–10595. doi: 10.18632/oncotarget.2513

Fig.4. AR enhances MMP9 expression by binding to its promoter region in SGC-7901 cells.

Fig.4

(a) MMP9 promoter region and AR-binding sites predicated by two different on-line tools. Bold on the left indicated the amplified portion of the promoter region, and bold on the right (Up and Low) indicated the binding sites with high scores. (b) Luciferase report assays. A reporter plasmid for MMP9 (pGL3-MMP9) was generated by cloning the MMP9 promoter region (wt) or its mutants (mut1 and mut2) into the pGL3-basic vector. AR significantly increased the luciferase activity of MMP9 promoter region, which was significantly reduced by knockdown of AR. Mut1 may contained the AR-binding site. (c) MMP9 promoter region and AR interaction validated by ChIP assays. MMP9 cDNA was detectable in the immunoprecipitated chromatin samples of SGC7901-AR cells using an antibody against AR, suggesting that AR binds to the MMP9 promoter. Genomic DNA and IgG were used as controls. Results are the mean of three independent experiments ± SD (*P<0.05).