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. 2007 Oct 12;407(Pt 3):461–469. doi: 10.1042/BJ20070658

Figure 4. Functional FXR-response elements are present within the fetuin-B P2 promoter.

Figure 4

(A) Human fetuin-B P2 promoter sequence was aligned with genomic sequences from different species. Two putative FXR-binding sites (FXRE1 and FXRE2) were identified in the human sequence within 1.6 kbp upstream of the predicted transcriptional start site. Boldface letters and arrows indicate putative IR-1 sites. (Mouse chromosome 16: position 19829761 to 19829857 bp; bovine chromosome 1: position 48748 to 48848 bp; Canis chromosome 34: position 6763848 to 6763947 bp.). HepG2 cells were transfected (B, C) with the reporter construct in the presence of either empty vector (control) or FXR and RXR expression vectors. Cells were treated with 50 μM CDCA for 36 h. RLU (relative light unit) signal was normalized to the β-galactosidase activity. FXR agonist-mediated induction is expressed as the fold change over DMSO. (B) Human fetuin-B P2 promoter (−1978 bp/+263 bp) was cloned into the pGL3-basic reporter vector. Mutations were introduced in the FXRE1 or FXRE2 elements (E1mt, E2mt) as described in the Materials and methods section. Values are means±S.E.M. (n=3). (C) HepG2 cells were transfected with reporter constructs containing three copies of either the FXRE1 or FXRE2 element. TK-pGL3 plasmid was used as a control. Mutations were introduced in the FXRE1 or FXRE2 elements (E1mt, E2mt) as described in the Materials and methods section. Luc, luciferase.