Gel mobility shift assay for the TMP21 gene promoter. Gel shift assays were performed as described in the Material and Methods. Double stranded NFAT oligonucleotide probes labeled wuth IRDye-680 were applied. Lane 1 is labeled probe alone without nuclear extract. Incubation with nuclear extracts retarded the migration rate of the labeled probe, forming a new shifted DNA-protein complex band (lane 2). Competition assays were performed by further adding different concentrations of molar excess of unlabeled competition oligonucleotides, consensus NFAT (lane 3, 4), binding sequence mutant NFAT (lane 5, 6) and TMP21 NFAT elements (lane 7, 8, 9, 10).