NF-Y binds the −4,425/−4,392 G6PC2 promoter region in vitro. NF-Y binds the −4,425/−4,392 G6PC2 promoter region in vitro. A: The sense strand sequences of the oligonucleotides used in gel retardation assays are shown. SNP base pairs are shown in bold lower case letters. The consensus NF-Y binding motif, CCAAT, is taken from Quandt et al. (31). B: βTC-3 nuclear extract was incubated in the absence or presence of the indicated anti-serum for 10 min on ice. A labeled oligonucleotide representing the −4,425/−4,392 G6PC2 promoter region and containing the rs13431652-A allele (rs13431652-A; panel A) was then added and incubation continued for 20 min at room temperature. Protein binding was then analyzed using the gel retardation assay as described in Research Design and Methods. In the representative audioradiograph shown, only the retarded complexes are visible and not the free probe, which was present in excess. Nabila Bouatia-Naji et al. Diabetes 2010;59:2662-2671 ©2010 by American Diabetes Association