FIG. 6.

Effect of the HNF4 inhibitor on the electrophoretic mobility of various HNF4 deletion constructs. Nuclear extracts (3 μl) from HEK 293 cells transfected with an expression vector encoding either Myc-tagged HNF4α (lanes 1, 2, 5, and 6), Myc-tagged HNF1α (lanes 3, 4, 7, and 8), Myc-tagged C-terminal deletion construct Mt-HNF4αΔEF (lanes 9 and 10), or the N-terminal deletion construct HNF4αΔAB (lanes 11 and 12) were subjected to electrophoresis on a native 4% polyacrylamide gel. Prior to electrophoresis, the extracts were mixed with 1 μl of MonoQ-purified HNF4 inhibitor as indicated. The gels were blotted and probed with the Myc-specific antibody 9E10 (lanes 1 to 10) or the HNF4α-specific monoclonal antibody H4/39f (32) (lanes 11 and 12). The gel with lanes 5 to 8 was denatured before blotting. Lanes 13 and 14 are blotted from an SDS–15% polyacrylamide gel containing Myc-tagged transfected HNF4α and probed with the Myc tag-specific antibody 9E10.