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. 2013 Nov;54(11):3030–3044. doi: 10.1194/jlr.M038323

Fig. 7.

Fig. 7.

Mutational analysis of IR1a and IR1b in BSEP promoters. Four human BSEP promoter mutants with mutation in IR1a, IR1b, the half ER2 site, or both IR1a and IR1b were made using human BSEP promoter reporter phBSEP(-2.6kb) as parental template, resulting in mutants phBSEP(-2.6kb)-IR1a-Mut, phBSEP(-2.6kb)-IR1b-Mut, phBSEP(-2.6kb)-IR1ab-Mut, and phBSEP(-2.6kb)-ER2-Mut. Four equivalent mutants were constructed using rat bsep promoter reporter prBSEP(-2kb) as parental template, resulting in prBSEP(-2kb)-IR1a-Mut, prBSEP(-2kb)-IR1b-Mut, prBSEP(-2kb)-IR1ab-Mut, and prBSEP(-2kb)-ER2-Mut. The four human BSEP promoter mutants and wt were cotransfected with (A) FXRα1 or (B) FXRα2 into Huh 7 cells. The four rat bsep promoter mutants and wt were cotransfected with (C) FXRα1 or (D) FXRα2. Sixteen hours posttransfection, cells were treated with DMSO (0.1%) or CDCA (10 μM) for 30 h, followed by detection of luciferase activities by the dual-luciferase reporter assay system. The data are presented as mean ± SD of at least three repeated experiments. *P < 0.05 between DMSO and CDCA-treated cells (Student t-test). The fold inductions by CDCA over DMSO are presented on the top of the bars.