FIG. 7.

Gel shift analysis reveals four transcription factor binding sites in region 4.2. Gel shift analyses with nuclear extract (NE) from primary human keratinocytes (hK) were performed by using overlapping oligonucleotides spanning the entire region 4.2. Oligonucleotides which bound specific complexes are shown. The identities of the proteins which bound are indicated at the left, and an asterisk (∗) denotes larger complexes (supershifted complexes) that formed with the addition of the antibodies (Ab) indicated. Additional details are outlined in the legend to Fig. 5. Shown are data indicating protein-DNA complexes for (A) Sp1/Sp3 and oligonucleotide A; (B) AP-2 family members and oligonucleotide B; (C) NF-I family members and oligonucleotide NF-I; and (D) G/C2 and oligonucleotide G/C2. NS, nonspecific complex. Competitor oligonucleotides: AP-2, consensus AP-2 binding site (Promega); AP-2m1, oligonucleotide containing the sequences in oligonucleotide B but with a mutated AP-2 site (CCC TGA GGG → CCC TGG AAT); G/C2m, GGG → AAA mutation as described in the text; NF-Im, a mutation in the core consensus sequence of NF-I (GCCAA → GTTAA). Antibodies used are described in Materials and Methods.