Figure 4.

HNF-1β is required for the expression of Kif12 in mIMCD3 cells and mouse kidney. (A) Schematic diagram of wild-type mouse HNF-1β (top) and the dominant negative HNF-1βΔC mutant. Boxes indicate Flag (red) and EGFP (green) epitope tags. (B) Expression of the HNF-1βΔC mutant was confirmed by immunoblot analysis of lysates from mIMCD3 cells induced with mifepristone (Mif). (C) The in vivo association of the HNF-1βΔC mutant with the Kif12 promoter was confirmed by ChIP assay using an anti-Flag antibody in transfected mIMCD3 cells (n = 3). (D) Induction of the HNF-1βΔC mutant in mIMCD3 cells resulted in decreased expression of endogenous Kif12 mRNA (n = 6). (E) Treatment of parental mIMCD3 cells with mifepristone had no effect on the expression of Kif12 mRNA. (F) Kif12 mRNA expression was decreased in cystic kidneys from P8 kidney-specific HNF-1β knockout mice compared with wild-type controls. Error bars indicate SD (n = 3).