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. Author manuscript; available in PMC: 2009 Feb 6.
Published in final edited form as: J Biol Chem. 2007 Jan 10;282(14):10742–10748. doi: 10.1074/jbc.M610997200

FIGURE 3. BMP-2 and Runx2 activate Smad6 promoter through the OSE2-a site.

FIGURE 3

a, diagram shows the construct harboring wild-type (wt) or mutant (mt) OSE2-a and OSE2-b sites derived from −1191/+45 reporter. The wt and mt sequences of OSE2-a and OSE2-b sites are indicated. b, C2C12 cells were transfected with wt and mt −1191/+45 Smad6 reporter with Runx2 expression plasmid or treated with BMP-2 (100 ng/ml). Cell lysates were collected 24 h after transfection, and a luciferase assay was performed. Mutation of OSE2-a caused a significant reduction in basal promoter activity and completely abolished BMP-2- or Runx2-induced promoter activity of the −1191/+45 reporter. Mutation of OSE2-b had no significant effect on the basal promoter activity or the responsiveness to Runx2 or BMP-2. *, p < 0.05, unpaired Student’s t test, compared with vector alone. c, C2C12 cells were transfected with wt and mt −2006/+45 Smad6 reporter with Runx2 or Smad1 expression plasmid or treated with BMP-2 (100 ng/ml). Mutation of OSE2-a in −2006/+45 reporter significantly reduced basal promoter activity and inhibited BMP-2, Runx2, or Smad1-induced promoter activity. *, p < 0.05, unpaired Student’s t test, compared with vector alone.