Electrophoretic mobility shift assay of BMPRE2/bZIP/HNF4α/COUP. A probe corresponding to the BMP-RE2/ bZIP/HNF-4/COUP region was 32P labeled and nuclear extracts from 2 independent preparations (numbered 1 and 2) of 129 SvJ liver nuclear extracts (A,B) or lysates from HNF4α or SMAD4 overexpressing HEK293T cells (293T) were added (C). Each mouse liver sample was prepared from a group of 4 mice on an iron-deficient diet. (A) Nuclear extracts were either preincubated with buffer (none) or DNA competitor corresponding to the consensus motif of HNF4α (HNF4). (B) Nuclear extracts were either preincubated with buffer (none) or DNA competitor corresponding to the consensus motif of SMAD 3/4. Panels A and B were part of the same gel but panel B required a longer exposure to visualize the SMAD bands. This experiment is representative of more than 3 experiments.
