Genetic and Functional Variants Analysis of the GATA6 Gene Promoter in Acute Myocardial Infarction

Figures

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  Figure 1. The DSVs and SNPs within the GATA6 gene promoter identified in AMI patients and controls. Schematic representation of the identified GATA6 gene DSVs. The DSVs were named according to their locations in the GATA6 genomic sequences (NCBI: NC_000018.10). The transcription starts at 22169437 in the first exon that is untranslated.

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  Figure 2. Sequencing chromatograms of the DSVs and SNPs within the GATA6 gene promoter. (A). Sequencing chromatograms of the DSV and SNPs in AMI patients. (B). Sequencing chromatograms of the DSV and SNP in healthy controls. All sequence orientations of the DSVs and SNPs are forward. Top panels show wild type and bottom panels heterozygous DSVs or SNPs, which are marked with arrows.

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  Figure 3. LD analysis of the six SNPs in AMI. Standard color schemes indicate different levels of LD. Bright red: LOD > 2, D' = 1; Purple: LOD < 2, D' < 1; White: LOD < 2, D' < 1; LOD: Log of the odds of there being LD between two loci.

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  Figure 4. Relative transcriptional activity of wild-type and variant GATA6 gene promoters. Wild-type and variant GATA6 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 GATA6 gene promoter was designed as 100%. Relative activities of GATA6 gene promoters were calculated. (A) Relative activities of wild-type and variant GATA6 gene promoters in HEK-293 cells. Lanes 1, pGL3-basic; 2, pGL3-WT; 3, pGL3-22168362A; 4, pGL3-22168409G; 5, pGL3-22168438A; 6, pGL3-22168521T+22168944A+22169265A; 7, pGL3-22168521T; 8, pGL3-22168780G. (B) Relative activities of wild-type and variant GATA6 gene promoters in H9c2 cells. Lanes 1, pGL3-basic; 2, pGL3-WT; 3, pGL3-22168362A; 4, pGL3-22168409G; 5, pGL3-22168438A; 6, pGL3-22168521T+22168944A+22169265A; 7, pGL3-22168521T; 8, pGL3-22168780G. WT, wild-type. *, P 0.05; **, P 0.01.

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  Figure 5. EMSA of biotin-labeled oligonucleotides containing DSV and SNP. Wild-type and variant oligonucleotides (31 bp) were designed and labeled with biotin for the DSV and SNP identified in AMI patients, including g.22168362C>A (rs1416421760) and g.22168409A>G. 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.