CBF-A binds differentially to CArG boxes from LAB and HAB lines. A, SNPs embedded in putative consensus DNA-binding sites (DBS) as predicted by the TESS program. Only those DBS are listed in which the respective transcription factor (TF) is known to be expressed in the PVN. B, Recombinant CBF-A protein binds specifically to the CArG element from the AVP promoter of the LAB line forming two complexes in an EMSA. The major low mobility and minor higher mobility complexes (thin arrows) are both supershifted (thick arrow) with increasing concentrations of GST antibody, as indicated at the top. C, Increasing concentrations of CBF-A reveal a higher affinity for the LAB (L) double-stranded CArG box than for the HAB (H) sequence in EMSA. D, The unlabeled LAB oligonucleotide competes more efficiently than the unlabeled HAB oligonucleotide for CBF-A binding to the LAB sequence, in EMSA, demonstrating that the CArG element from the LAB animal binds CBF-A more efficiently. The results in B-D are representative of three individual experiments.