Binding of Ets transcription factors to the TM promoter. HUVECs were stimulated with or without 80 ng/mL human TNF-α for 16 hours. (A) Representative EMSA performed with primers/probe containing the sequence of the TNF-α response element within the TM promoter. Preincubation of HUVEC nuclear extracts with unlabeled oligonucleotides containing Ets binding sequences from the stromelysin promoter (EtsSTR), but not a nonspecific (NS) oligonucleotide containing the potential noncanonical NF-κB binding site, competitively reduced the observed the intensity of the dominant DNA-protein complex (solid arrow). (B) Chromatin immunoprecipitation was performed by using a probe specific for the TM promoter and antibodies specific for various Ets species. Specificity for the TNF-α response element within the TM promoter was confirmed by lack of enrichment above background by using primers and probe to a sequence approximately 18 kb upstream of the TM promoter (data not shown). ▪ indicates control; ▦, stimulation with TNF-α. Values are the mean ± SEM for 4 experiments. P values were not significant for paired vehicle and TNF-α groups.