FIGURE 4.

The CRE-like site is functionally important for basal and TGFβ induction of Csrp2 promoter activity. A, conservation of the putative CRE site among species. Sequence alignment of corresponding regions of human and rat promoter sequences to the mouse promoter. A putative CRE site (TAACGTCA) is in boldface type and underlined in the mouse sequence. B, the putative CRE site is important for Csrp2 promoter activity. The –795 Csrp2 wild type (wt) and CRE mutant (mut) promoter constructs are schematically depicted in the left panel. VSMCs were transiently transfected with Csrp2 luciferase reporter constructs (500 ng/well) containing –795 or –795CREmut with putative CRE site mutated and pCMVβ (500 ng/well) to normalize for transfection efficiency in triplicate using FuGENE 6 transfection reagent. Two hours after transfection, cells were treated with or without TGFβ (10 ng/ml) for 24 h. Cells were then harvested for luciferase and β-galactosidase activity assays. Luciferase activity is expressed relative to –795 without TGFβ treatment.