4.

In vitro binding of HNF1 to 7/8 enhancer. EMSA using Caco2 nuclear extracts (A) or in vitro translated HNF1 proteins (B). (A) Two major protein complexes designated A and B with solid arrowheads are formed with a radiolabelled oligo -nucleotide spanning the 85 + 10 kb DHS region. Competition with 100-fold molar excess of unlabelled wild-type (100 × self) and mutant (100 × mut) probes is indicated. HNF1-specific antibodies (to either α or β) were used in supershift analysis, and free radiolabelled oligonucleotide (probe) and goat IgG were used as controls. Dashed arrow indicates HNF1α supershifted band in panel A; indicates non-specific band detected in HNF1α lane.