Figure 4.

In Vitro Binding of LeCp to Promoter Sequences of Acs2.

Gel-shift binding assay was performed by incubating LeCp (0.1 μg) with 30 fmol of end-labeled 40-bp fragment (corresponding to −715 to −675 in the Acs2 promoter) and 2-, 10-, 25-, 50-, or 250-fold excess competitor fragments, as indicated (left). LeCp (0.1 μg) was also incubated with 30 fmol of end-labeled wild-type (W.T.) or mutated 40-bp fragment (corresponding to −715 to −675 in the Acs2 promoter), as indicated, in the presence of 2 μg of poly(dI-dC) (right). After the binding step, the DNA–protein complexes were fractionated by PAGE and detected by autoradiography. Bound complexes are identified at left. oligo, oligonucleotide.