FIGURE 6.

Representative EMSA to identify a partial consensus sequence for the ZTRE. A, an IRD-labeled probe (50 fmol), corresponding to the region −156 to +46 of the SLC30A5 gene, was electrophoresed through a non-denaturing 5% polyacrylamide gel after incubation with 5 μg of nuclear extract from cells treated with 150 μm zinc and in the absence or presence of specific, double-stranded oligonucleotides (200-fold excess), as indicated. The different oligonucleotides included different modifications to the ZTRE sequence, made as single substitutions at both halves of the palindromic sequence at the positions indicated. The arrow indicates the position of the major band representing probe bound to proteins in the cell extracts. B, intensity of the major band for the 150 μm zinc concentration (as in A; determined using UviPhotoMW image analysis software) in the presence of the oligonucleotide competitors, as indicated, was expressed as a percentage of the corresponding band in the absence of competitor and plotted as mean ± S.E. (error bars). n = 18 for oligonucleotide −124 → −75; n = 4 for all other oligonucleotides. ***, p < 0.001 by one-way analysis of variance followed by Bonferroni's multiple comparison test.