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. Author manuscript; available in PMC: 2013 Jul 16.
Published in final edited form as: Bone. 2012 Sep 29;52(1):145–156. doi: 10.1016/j.bone.2012.09.029

Fig. 6.

Fig. 6

Transactivation of BMP2 promoter by Wnt/β-catenin signaling through Tcf/Lef response elements. (A and B) Effects of inhibitor of interaction of β-catenin and TCF4 on BMP2 promoter activity. C2C12 cells were co-transfected with −2712/+165-Luc reporter and expression vectors for β-catenin and TCF4, and expression vectors for ICAT (A) or ΔTCF4 (B) for 36 h. Relative luciferase activity in the cell lysates was measured and normalized by β-gal activity. *: P<0.05 ICAT or ΔTCF4 vs. vector; #: P<0.05 ICAT+β-catenin/TCF4 or ΔTCF4+β-catenin/ TCF4 vs. vector+β-catenin/TCF4 (n=6). (C) Mutagenesis of TREs in the BMP2 promoter. The core nucleotides (bold dashes) of putative TREs at −2269/−2263 and −1824/−1804 (bold) in the mouse BMP2 promoter reporter −2712/+165-Luc were deleted using synthesized mutagenesis DNA oligonucleotides. (D) Effects of mutated TREs on β-catenin/TCF4 activation of BMP2 promoter activity. C2C12 cells were co-transfected with−2712/+165-Luc reporter containing wild-type TREs or mutated TREs (Δ-2269/−2263, Δ-1824/−1804) with expression vectors for β-catenin and TCF4 for 36 h. Relative luciferase activity in the cell lysates was measured and normalized by β-gal activity.*:P<.01 β-catenin/TCF4 vs. vector (n=6).