![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=1722502287&Signature=sgmWvX7-nha3NTe2fIY3fSPJ48ZwLnzBfnYuyKSKPL1E0UNelyJXuP2ED~1HoC4puVhUMOaBsuFSTKAqYK~laN-6Ldny5~QAMGwhj4HiYrnL8u0KwRqOhKFxMq~Y4QulnlXqUQ6-8VApI~S8oi1NxH04tCnBtqtiw6ihSOC5ySh8Fi4Og6sGLCwizA8Renu1A5S3PioqMRVt1U0-CzBzCQrA6kMfi1MI-XCIbDTkf1tvD0y2m3I5op67dytSf3deNdjMv-luWJ8EAqVVB0DnP1BcmU9HOpE7Zr3Oczm92OmrDe1aMiRr2gDKr33L1W8tz9rXePMaqOSjGl300Qo2DQ__&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).
