Transcriptional regulation of human α1(I) procollagen gene in dermal fibroblasts

Figure 4 Result of EMSA with EcoR II-digested -268 to +42 bp as probe. Lane1: Labeled DNA, Lane2: Labeled DNA+ nuclear protein +NF-1 consensus DNA, Lane3: Labeled DNA + nuclear protein + Sp-1 consensus DNA, Lane4: Labeled DNA + nuclear protein + Ap-1 consensus DNA, Lane 5: Labeled DNA + nuclear protein, Lane 6: Labeled DNA + excess unlabeled DNA + nuclear protein. The existence of several retarded bands in EMSA indi-cated that there were several nuclear protein binding sites in sequence –268 to +42 bp (lane 5). No retardation occurred when excess unlabeled DNA probe was added to the DNA-protein reaction, confirming the specificity of the retardation (lane 6). Consensus DNAs for Sp1, Ap-1 and NF-1 were among the pos-sible regulatory elements since the molar excess of the consen-sus unlabeled probe (Sp1, Ap-1, NF-1) inhibited partially the formation of retardation bands differently (lanes 2, 3 and 4).