Binding site specificity of Stat6 decoy.

(A) D10 cells were incubated with medium (−) or 100 nmol/L IL-4 (+) for 10 minutes at 37°C. Nuclear extracts were incubated with a [³²P]-labeled Stat6 oligonucleotide probe corresponding to the Cε gene promoter. Competition was performed with 100-fold molar excess of unlabeled Stat6 decoy ODN (lane c), scrambled ODN (lane d), or cold Cε element ODN (lane e). Arrow indicates migrational location of Stat6-DNA complex or free probe. (B) D10 cells were incubated with medium (−) or 100 nmol/L IL-2 (+) for 10 minutes at 37°C. Nuclear extracts were incubated with a [³²P]-labeled Stat5 oligonucleotide probe corresponding to the prolactin response element of the β-casein gene promoter. Competition was performed with 100-fold molar excess of unlabeled cold Stat5 ODN (lane c), Stat6 decoy ODN (lane d), or scrambled ODN (lane e). Arrow indicates migrational location of Stat5-DNA complex or free probe. (C) D10 cells were incubated with medium (−) or 100 nmol/L IL-2 (+) for 10 minutes at 37°C. Nuclear extracts were incubated with a [³²P]-labeled Stat1/3 oligonucleotide probe corresponding to the SIE gene promoter. Competition was performed with 100-fold molar excess of unlabeled cold Stat1/3 ODN (lane c), Stat6 decoy ODN (lane d), or scrambled ODN (lane e). Arrow indicates migrational location of each Stat1- or Stat3-DNA complex or free probe.