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Blood, Vol. 112, Issue 6, 2336-2339, September 15, 2008
A GABP-binding element in the Robo4 promoter is necessary for endothelial expression in vivo
Blood Okada et al.
112: 2336
Supplemental materials for: Okada et al
Files in this Data Supplement:
- Figure S1. GABP selectively binds to the −119 ETS (1) site of the human Robo4 promoter (JPG, 151 KB)
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(A) Nucleotide sequence and alignment of the 5′-flanking region of the human, mouse, and rat Robo4 genes shows 4 conserved ETS consensus motifs. Arrow represents start site of transcription. ETS(1) represent the GABP-binding -119 ETS motif. (B) Transient transfection of HCAEC with pGL3 containing the wild-type 3-kb Robo4 promoter (WT) or the 3-kb Robo4 promoter with mutations of ETS(1), ETS(2), ETS(3) or ETS(4). Luciferase light units are corrected for transfection efficiency. Data represent mean +/− SE of 14 replicates. * p< 0.05 between WT and mutant promoters. (C) EMSA was performed with 32P-labeled probe spanning each of the putative ETS sites (or ETS(1) mut) in the absence (lanes 1, 3, 5, 7, 9) or presence of nuclear extract from HCAEC (lanes 2, 4, 6, 8, 10). The bracket indicates GABP-DNA complex. NS denotes non-specific band.
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