Figure 5
From: DEK binding to class II MHC Y-box sequences is gene- and allele-specific
Recombinant DEK protein (rDEK) binds in a sequence-specific manner to the DQA1*0101 Y box, but not to the DRA (consensus) Y box. (a) 5 μl of antisense rDEK (lane 2), 5 μg of Namalwa cell nuclear extract (lane 3), or 5 μl of partially purified recombinant DEK protein (rDEK) (lanes 4–12) was used in each EMSA binding reaction; rDEK was preincubated with the indicated unlabelled competitor before addition of the DQA1*0101 Y-box probe. Lane 1 contains no protein. (b) The indicated unlabelled oligonucleotide competitors (20 ng) were added to the binding reaction containing 5 μl of partially purified rDEK before addition of radiolabeled DQA1*0501 probe (lanes 1–4), or radiolabeled DRA probe (lanes 5–8). PolyD(I-C) (10 ng) was added to each binding reaction as nonspecific competitor. EMSA = electrophoretic mobility shift assay.