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. 2003 Jun 1;31(11):2769–2777. doi: 10.1093/nar/gkg396

Figure 4.

Figure 4

PKNOX1 is present in SH-SY5Y nuclear extracts and binds specifically to the Pbx/POU binding site of the FABP7 promoter. (A) Specific competitions (100-, 1000- and 10 000-fold molar excess) with cold wt probe and non-specific competition with the GRE oligonucleotide (10 000-fold molar excess). (B) Specific competitions with cold mutated del probe (100-, 1000- and 10 000-fold molar excess). (C) Disruption of the PKNOX1-binding complexes by co-incubation with an anti-PKNOX1 antibody. The asterisk on lane 5 indicates that the antibody was added from the beginning of the reaction instead of addition after the 10 min incubation on ice (see Materials and Methods). (D) PKNOX1 binds specifically to the Pbx/POU target site and this binding is abolished by the anti-PKNOX1 antibody. The presence of nuclear extract, purified recombinant PKNOX1 (10 ng), type of oligonucleotide probes, competitors and antibodies used are indicated above each lane. Specific retarded bands due to protein binding at the Pbx/POU target site are indicated by arrows. Note their disappearance after addition of the anti-PKNOX1 antibody.