Genetic variants of VEGFR-1 gene promoter in acute myocardial infarction

Figures

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  Fig. 1. Locations of the identified DSVs and SNPs within the VEGFR-1 gene promoter and 5′-UTR. The genetic variants are named according to the genomic DNA sequence of the human VEGFR-1 gene (Genbank accession number NC_000013.11). The transcription start site is at the position of 28495128 in the first exon

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  Fig. 2. Sequencing chromatograms of the DSVs and SNPs. a Sequencing chromatograms of the DSVs and SNPs only identified in AMI patients, g.28495805C>T (rs111458691), g.28495796 G>T, g.28495445 G>T, g.28495341 A>G, g.28495138 G>C, and g.28494900 G>A. b Sequencing chromatograms of the DSVs identified only in controls, g.28495678 G>A and g.28495356G>A. Top panels show wild-type and bottom panels heterozygous sequences, which are marked with arrows

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  Fig. 3. Relative transcriptional activity of wild type and variant VEGFR-1 gene promoters. Wild-type and variant VEGFR-1 gene promoters were cloned into reporter gene vector pGL3 and transfected into cultured cells. The transfected cells were collected, and dual-luciferase activities were assayed. Empty vector pGL3-basic was used as a negative control. Transcriptional activity of the wild-type VEGFR-1 gene promoter was designed as 100%. Relative activities of variant VEGFR-1 gene promoters were calculated. a Relative activities of wild type and variant VEGFR-1 gene promoters in HEK-293 cells. b Relative activities of wild-type and variant VEGFR-1 gene promoters in H9c2 cells. Lanes 1, pGL3-basic; 2, pGL3-WT; 3, pGL3-28494900A; 4, pGL3-28495138C; 5, pGL3-28495341G; 6, pGL3-28,495,445 T; 7, pGL3-28,495,796 T; 8, pGL3-28,495,805 T; 9, pGL3-28495356A; and 10, pGL3-28495678A. WT wild type, AMI AMI patients, CTR healthy controls. *P < 0.05, compared to pGL3-WT; **P < 0.01, compared to PGL3-WT

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  Fig. 4. EMSA of biotin-labeled oligonucleotides. Wild-type and variant oligonucleotides (30 bp) were designed and labeled with biotin for genetic variants identified in AMI patients, including g.28495805C>T(rs111458691), g.28495138G>C, and g.28494900G>A. EMSA was conducted with biotinylated oligonucleotides and the nuclear extracts from HEK-293 and H9c2 cells. Free probe was marked with an arrow at the bottom. The affected binding for transcription factors was marked with an open arrow