Supershift assay of the Pax4 are shown.

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Supershift assay of the Pax4 are shown. Supershift assay of the Pax4 are shown. An equal amount of protein yielded by the ivTT system was applied in each lane. The mouse wild-type Pax4 (lanes 4, 5, and 6) and mouse mutant Pax4 (R129W) (lanes 7 and 8) were used as His-tagged fusion protein. Mouse R129W corresponds to human R121W. The anti-HisG antibodies were added in lanes 3, 5, and 8. Mouse serum was added in lane 6. Unprogrammed wheat germ cell extract (lane 1) and extract derived from nonrecombinant plasmid [pcDNA3.1(His)] (lanes 2 and 3) were applied as negative controls. The long arrow indicates specific binding of Pax4 with His-tagged fusion protein to the probe. The short arrows indicate nonspecific binding. The oligonucleotide probe was rat glucagon promoter G3 element (GluG3). Identical results were obtained in three independent experiments. Yoshinori Shimajiri et al. Diabetes 2001;50:2864-2869 ©2001 by American Diabetes Association