Fig. 5
The −1690 bp substitution in the Meishan promoter binds GATA-4, whereas the Control/Index promoters bind NF-κB. a Oligonucleotide probes (5’ to 3’) were synthesized containing sequence flanking a naturally occurring point mutation at −1690 of the Control/Index (C/I −1700/−1680) or −1666 of the Meishan (M −1676/−1656) promoter. Underlines within each DNA probe represent the nucleotide substitution between lines and putative NF-κB (ACCCCA; Blue) and GATA-4 (ATATCT; Red) elements identified by sequence analysis are highlighted. b EMSAs were performed with αT3-1 nuclear extracts and radiolabeled oligonucleotides spanning the −1690 bp substitution between the Control/Index and Meishan (−1666 bp) swine lines. To determine the specificity of the DNA:protein complex, 50-fold molar excess of either unlabeled homologous or heterologous DNA were added (specific complexes indicated by arrows). c To assess which specific proteins comprised the DNA:protein complex binding to the oligonucleotide spanning the substitution at −1690 bp of the Control promoter (−1666 bp of the Meishan promoter), antibodies directed against the p65, p50, and p52 subunits of NF-κB, GATA-1, -2 and -4, or an equal mass of rabbit IgG were added (supershifts indicated by arrows)