FIGURE 3.

Egfl7 expression in endothelial cells is regulated by TNFα at the transcriptional level. A, Egfl7 transcript levels measured by duplex RT-qPCR in confluent HUVEC treated with 10 ng/ml TNFα (●) or with PBS (□) for 1 h and then with 10 μg/ml actinomycin D (ActD, t = 0) and assessed during the next 6 h. B, Egfl7 transcripts levels measured by duplex RT-qPCR in confluent HUVEC treated with DMSO or 10 μg/ml actinomycin D for 1 h before stimulation with or without 10 ng/ml TNFα for 6 h. *, p < 0.05; **, ns, non-significant. C, luciferase activities measured in HUVEC transfected with pGL3basic (Ctrl) or with the indicated reporter constructs containing fragments of the human egfl7 gene promoter and with the pCMV-β-Gal normalizing vector. The cells were then treated with 10 ng/ml TNFα (black bars) or with PBS (white bars) and lysed 18 h later. The letters correspond to conserved promoter regions (16). The numbers indicate the base position relative to the exon 1b transcription initiation site (2). Activities were normalized with β-galactosidase values, folds of induction were calculated using pGL3basic values as reference; the results are representative of three experiments performed in triplicate. **, p < 0.01; ***, p < 0.001; ns, non-significant.