FIG. 7.
Mapping the OBP-2-binding site. (A) Oligonucleotides that span the 50-bp 7–2 oligonucleotide that contains the OBP-2 site were tested for their ability to be directly bound by OBPH7. For each oligonucleotide target, lane 1 contains a binding reaction mixture without any reticulocyte lysate, lane 2 contains reverse negative control lysate, and lane 3 contains lysate with truncated OBPH7 protein. (B) Competition EMSA with unlabeled oligonucleotides at 16- and 80-fold molar excess. −, no competitor was added. NP and R are negative controls as described in the Fig. 5 legend. In panels A and B, arrows point to the specific shifts. (C) Schematic diagram of oligonucleotides used and summary of binding and competition assay results shown in panels A and B. NT, not tested. ++ indicates a stronger degree of interaction of an oligonucleotide with OBPH7 than +; −, no competition observed.