![Fig. 9. PU.1 binds the predicted site in the MBP-P2 promoter. / COS-1 cells were transfected with empty (−) or PU.1 (+) expression vector, and total cell extracts were prepared 24 hours after transfection. Wild-type consensus PU.1 (left and middle panels; 5′-gggctgcttgaggaagtataagaat-3′) or wild-type MBP PU.1 (right panel; 5′-tctccctgggggaagttcctccaaggcc-3′) double-stranded oligonucleotides were end-labeled with [32P]-γ adenosine triphosphate (ATP). Labeled oligonucleotides (0.02 pmol) were incubated with 5 μg total cellular extract in the presence or absence of anti-PU.1 (1 μg) antibody (left and right panels) or unlabeled (0.2 pmol) wild-type or mutant oligonucleotides (consensus, 5′-gggctgcttgagagagtataagaat-3′; MBP PU.1, 5′-tctccctgggcaaagtttgtccaaggcc-3′). Arrows at the right of each panel indicate the positions of the complexes containing PU.1. Wt indicates wild-type; Mt, mutant; CON, consensus PU.1 binding site; MBP, major basic protein promoter PU.1 binding site; Ab, anti-PU.1 antibody (sc-352; Santa Cruz Biotechnology).](https://ash.silverchair-cdn.com/ash/content_public/journal/blood/101/8/10.1182_blood-2002-04-1039/4/h80834157009.jpeg?Expires=1735547636&Signature=sh5PaUW9aB71oKCOhWlYj7xbjWGcboGRdy99gyoo-~i-54zUW-yvprNpZ8ab9~deTSDHIiv9P68rFzr-tlITRNMy6AOUIgIshFCNnSdyRDqAWz700DWOSA-bLLtWoEgspOiGZqvtkmOGLFE3ztuHFjZFxZuQDkZp3kFWeQ1N4~FEkeqJs5FIXmx3jfF~3jzUl-wLDiVIs4DAWVXZE-fuHtM0asDoVy1cAqBN~lkTMhUEROjD3qwHP2fXM~KFn~1j1k6QkuOKrTvdcMDSiQArW6SWpAfQXJqGCYueokQPOvTBpW-4fT8dKmVhkKOVWd~-rMUrMwdEiCgFmakzmD6p7A__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
PU.1 binds the predicted site in the MBP-P2 promoter.
COS-1 cells were transfected with empty (−) or PU.1 (+) expression vector, and total cell extracts were prepared 24 hours after transfection. Wild-type consensus PU.1 (left and middle panels; 5′-gggctgcttgaggaagtataagaat-3′) or wild-type MBP PU.1 (right panel; 5′-tctccctgggggaagttcctccaaggcc-3′) double-stranded oligonucleotides were end-labeled with [32P]-γ adenosine triphosphate (ATP). Labeled oligonucleotides (0.02 pmol) were incubated with 5 μg total cellular extract in the presence or absence of anti-PU.1 (1 μg) antibody (left and right panels) or unlabeled (0.2 pmol) wild-type or mutant oligonucleotides (consensus, 5′-gggctgcttgagagagtataagaat-3′; MBP PU.1, 5′-tctccctgggcaaagtttgtccaaggcc-3′). Arrows at the right of each panel indicate the positions of the complexes containing PU.1. Wt indicates wild-type; Mt, mutant; CON, consensus PU.1 binding site; MBP, major basic protein promoter PU.1 binding site; Ab, anti-PU.1 antibody (sc-352; Santa Cruz Biotechnology).
