Fig. 2. HGF/SF neither changes relative DNA-binding activity of Sp1 and Sp3 factor nor induces additional Sp1-indendent binding to the -88/-65 bp VEGF/VPF promoter sequence. A representative EMSA using nuclear extracts of untreated (lanes 1, 3, 5, 7 and 9) or HGF/SF-stimulated HaCaT cells (10 minutes; lanes 2, 4, 6, 8 and 10). Unlabeled Sp1 consensus oligonucleotides (Promega) were used at a final concentration of 0.35 µM (lanes 3 and 4). Supershift analysis was performed by addition of specific Sp1 or/and Sp3 antibodies (Santa Cruz) at a final concentration of 100 ng/µl (lanes 5 to 10). The DNA sequence of the utilized probe is shown at the top (Sp1 sites underlined), the formation of Sp1-dependent binding complexes is indicated by arrows to the left; supershifted complexes are marked by a bold arrow to the right.