FIG. 2.
Gel shift assay of sites 2 and 3. A 0.1-pmol portion of radiolabeled PCR fragment containing sites 2 and 3 from the ACC1 ORF was used. C1, C2, and C3 represent one, two, and three Gal4 DBD dimers bound to the probe, respectively. (A) Lanes 1 to 6, 0, 0.0005, 0.0024, 0.012, 0.06, and 0.3 pmol of purified Gal4DBD(1–147). (B) For lanes 2 to 8, 0.15 pmol of Gal4DBD(1–147) was used in binding reactions. For competition, 0.01, 0.1, and 1.0 pmol of cold consensus UASGAL were added (lanes 3 to 5). Then 0.01, 0.1, and 1.0 pmol of cold mutated Gal4 binding sites were added to (lanes 6 to 8).