FIGURE 6.

TGF-β1 induces FXII promoter activity via SBE at position −272 to −269 (SBE-(−272/−269)) in a JNK-dependent fashion. A, schematic representation of FXII promoter luciferase reporter constructs. The angled arrow indicates the transcription start site. B, NIH3T3 cells were transfected with the indicated FXII promoter deletion constructs and then were either not stimulated (open columns) or were stimulated with TGF-β1 (closed columns). Luciferase activity is expressed in relative luminescence units (RLU). Data represent mean values ± S.D. from four independent experiments, each performed in triplicate; **, p < 0.01. C, schematic representation of the FXII promoter region containing a putative SBE at position −272 to −269. pGL3-299 C/T represents a construct in which the SBE-(−272/−269) was mutated by the replacement of the C residue at position −272 by T. D, NIH3T3 cells were transfected with the pGL3-299 or pGL3-299 C/T constructs. Luciferase activity was determined in untreated (open columns) and TGF-β1 treated cells (closed columns). Data represent mean values ± S.D. from four independent experiments, each performed in triplicate; **, p < 0.01. E, schematic representation of the pGL3-299 construct and the construct lacking SBE-(−272/−269) (pGL3-183ΔSBE-(−272/−269)). F, NIH3T3 cells were transfected with pGL3-299 or pGL3-183ΔSBE-(−272/−269), and the luciferase activity was measured in unstimulated (open columns) and TGF-β1-stimulated cells (closed columns). Data represent mean values ± S.D. from four independent experiments, each performed in triplicate; **, p < 0.01. G, NIH3T3 cells, transfected with the pGL3-299 construct, were pretreated with SB431542, SP600125, wortmannin (Wort), PD98059, or SB203580 for 1 h prior to incubation with TGF-β1. Luciferase activity was determined in untreated (open columns) and TGF-β1 treated cells (closed columns). Data represent mean values ± S.D. from four independent experiments, each performed in triplicate; *, p < 0.05; **, p < 0.01; ***, p < 0.001.