Fig. 4.

PXR represses FoxA2 trans-activation of the mouse Hmgcs2 promoter in Huh7 cells. A. Reporter plasmid, pGL3/HMGCS2 -3.0k was co-transfected with or without FoxA2 in the presence or absence of mPXR and hRXR as indicated. Cell treatments and data analyses were done as in Fig 2A. A series of pGL3/HMGCS2 reporter plasmids as indicated were co-transfected with or without FoxA2 for 48 h. Numbers on the vertical ax indicate lengths of the Hmgcs2 promoter cloned into the reporter plasmids. Relative luciferase activities were calculated by taking the activity of the cells transfected with the pGL3/HMGCS2 -3.0k without FoxA2 as one. C. In vitro-translated FoxA2 was incubated with ³²P-labeled double-stranded putative FoxA2 binding sites, H1 and H2 within the Hmgcs2 promoter and a FoxA2 binding site of IGFBP-1 promoter. Gel shift analyses were performed as in Fig 2C. D. Reporter plasmid, pGL3/HMGCS2 -3.0k and the deletion mutants of the putative FoxA2 binding sites, H1 and H2, were co-transfected with or without FoxA2 for 48 h. Relative luciferase activities were calculated by taking the activity of the cells transfected with the pGL3/HMGCS2 -3.0k without FoxA2 as one. Bars indicate mean ± standard deviations in A, B, and D.