Figure 1.

Study overview. A, Infographic to describe the 4 components of the study (i) in a population-based analysis (n=3205), the observational association between smoking, DNA hypomethylation of F2RL3 and increased risk of myocardial infarction was quantified, focusing specifically on myocardial infarction. (ii) The impact of differential DNA methylation at F2RL3 on platelet function and reactivity in humans was tested in a recall study. (iii) The effect of exposure to cigarette smoke extract on F2RL3 DNA methylation, mRNA expression and protein abundance in acute megakaryocytic leukemia (CMK) cells was tested experimentally. (iv) An exploration of the functional regulation of F2RL3 was conducted using a series of reporter constructs. B, UCSC browser view of the F2RL3 exon 2 region containing the 4 cytosine–phosphate–guanine (CpG) sites assayed (chr19: 16 889 744-16 889 857 bp). Custom track study-specific features (black) shows the 4 DNA methylation sites CpG_1 to CpG_4 that were assessed by pyrosequencing, rs773902 and a CCAAT binding factor recognition sequence. CpG_3 corresponds to the CpG labeled cg03636183 on the Illumina Infinium Human Methylation450 BeadChip (450 K) array. Selected epigenetic annotations integrated from BLUEPRINT based on data from a megakaryocyte cell lineage. Figure produced using UCSC Genome Browser⁵⁸ based on Genome Reference Consortium Human Build 38 (GRCh38/hg38). For a full gene view and additional epigenetic annotations see Figure S10A.