EMSA results.

(A) Labeled WT and Mut oligos of the second binding site (+113/+122) on the 5’ UTR are shown in lane 1 and 3. In the presence of Elk1-db, clear complex formation of Elk1-db and WT probe is observed (lane 2). Also, there is no shift in the reaction in which Mut probe is incubated with Elk1-db (lane 4) and competition (lane 5), as expected. (B) Either WT or Mut oligonucleotides of the third putative binding site (+253/+262) could not participate in any complex formation with Elk1-db protein (lane 2 and 4, respectively). Lane 1 and 3 represent WT and Mut probes both of which lack Elk1-db, and positive control reaction is indicated in lane 6. (C) Biotin-labeled WT and Mut probes of the predicted promoter binding site (-428/-419) are loaded as controls (lane 1 and 3, respectively). When Elk1-db protein is incubated with both probes, no shift is observed with either WT probe (lane 2) or Mut probe (lane 3) and in competition reaction (lane 5) which included 1000-fold molar excess of unlabeled WT probe. Positive control reaction in which the complex formation is observed is indicated in lane 6.