FIGURE 5.

VEGF-A¹⁶⁵ induced the formation of a transcriptional complex with M-CAT cis-element in HUVEC. HUVEC were stimulated with VEGF-A¹⁶⁵ for 0, 10′, 15′, 30′, and 60′. Nuclear extracts were used for EMSA assay using a biotin-labeled DNA oligo probe corresponding to the M-CAT site of the DSCR1-1L promoter. Specific DNA-protein complex induced by VEGF-A¹⁶⁵ was indicated by a long arrow and nonspecific shifted band by a short arrow. 10-fold and 50-fold excess of unlabeled wild-type (lanes 7 and 8) or mutant probes (lanes 9 and 10) were included in the reaction mixtures with nuclear extracts from HUVEC stimulated by VEGF-A¹⁶⁵ for 15 min. The VEGF-A¹⁶⁵-induced shifted band was titrated off by an excess of unlabeled wild-type probes, but not unlabeled mutant probes. The data represent three independent experiments.