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. 2006 Nov 26;34(20):5951–5965. doi: 10.1093/nar/gkl689

Figure 5.

Figure 5

Dlx2 and LEF-1 synergistically activate the Msx2 promoter. (A) Schematic of the Msx2 promoter constructs used in transient transfection assays showing the location of Dlx2/Msx2 shared binding sites by asterisks. Note that the Msx2-238 Luc minimal promoter does not contain a Dlx2/Msx2 binding element. (B) CHO cells were transfected with the Msx2-5820, Msx2-872 or Msx2-238 luciferase reporter gene (5 μg). The cells were co-transfected with the CMV-Dlx2 and/or CMV-Lef-1 ΔN113 short isoform expression plasmids or the CMV plasmid without Dlx2 or Lef-1 (−) (2.5 μg). (C) C3H10T1/2 cells were transfected as in (B) to determine if the activation was cell dependent. To control for transfection efficiency, all transfections included the SV-40 β-galactosidase reporter (0.5 μg). Cells were incubated for 24 h and then assayed for luciferase and β-galactosidase activities. The activities are shown as mean fold activation compared to the Msx2 promoter plasmids without Dlx2 or Lef-1 expression and normalized to β-galactosidase activity (+/− SEM from eight independent experiments for (B) and from five experiments in (C). The mean Msx2 promoter luciferase activity with Dlx2 expression was ∼150 000 light units per 15 μg protein and the β-galactosidase activity was ∼75 000 light units per 15 μg protein.