ZPT2-14 binds to two core sequences separated by a particular spacing. (a), Distribution of spacing between two AGT/ACTs in the ZPT2-14-binding sequences selected by SAAB imprint assay (in AGT-AGT, AGT-ACT, and ACT-AGT configurations). Total of the three configurations are also shown. (b, c), Competition analysis of the ZPT2-14-binding sequences in gel mobility shift assays. ³²P-end-labeled probes with 5-base spacing—ACT-5-AGT (b)—or 10-base spacing—AGT-10-AGT (c)—were incubated with ZPT2-14ZF in the presence or absence of double-stranded competitors of ACT-5-AGT or GCG-5-GGC sequences in standard reaction mixtures. The amounts of competitors used were 0.2 ng (×1), 2.0 ng (×10), and 20 ng (×100). Central sequences of the probes and competitors are shown under the panels, with putative core sites underlined. Sequences of the probe DNA flanking the central regions are 5CTCGAGAAATTTCCCGGAATT-central region-CCGGAATTTCCCGGGGGGATCC-3′. The competition experiments were done twice and gave similar results.