BIRC6 Is Associated with Vulnerability of Carotid Atherosclerotic Plaque

Figure A1. Proxies of rs35286811. Plot represents individual LD r2 values of SNPs (ochre circles) in the BIRC6 region with rs35286811 (blue circle). GWAS-derived SNPs rs72798738 and rs11678584 are indicated. Regulatory potential of individual SNPs is scored from 1 (high) to 7 (low). Genes located in this interval are depicted in the bottom part of the figure. Image information was produced by means of https://ldlink.nci.nih.gov/.

Figure 1. BIRC6 gene structure. Known BIRC6 isoforms and their exon composition are indicated based on the ENSEMBL database, with flags on the right indicating transcript support level (TSL; TSL1 is the highest score where all splice junctions of the transcript are supported by at least one non-suspect mRNA). SNP rs35286811 is located in exon 67. BIRC6 gene expression primers (Hs.PT.58.21125813) were designed to amplify a fragment of mRNA encompassing part of exon 50 and exon 51, which is unique to the full-length isoform BIRC6-201. Average gene expression counts detected by RNA-Seq in vascular smooth muscle cells (VSMC) extracted from carotid plaque for BIRC6-201 and BIRC6-203 are indicated on the right side in bold; other BIRC6 isoforms were not detected in these cells [20]. 1 Bp: base pairs. 2 Flags indicate the transcript support level (TSL) of each isoform. 3 Avg GE: average gene expression counts obtained from RNA-Seq data.

Figure 2. BIRC6 gene expression levels (A) by RNA-Seq in the original VSMC samples extracted from 14 patients [20], and (B) by qPCR in extracts of carotid plaques from 70 asymptomatic and 104 symptomatic patients normalized for GAPDH and β-actin levels. In (C), BIRC6 gene expression levels from (B) are shown classified according to rs35286811 genotype.

Figure 3. RDW-CV values in (A) symptomatic and asymptomatic patients, and (B) following stratification for rs35286811 genotype (n = 285). (C) MAP1LC3B gene expression levels by qPCR normalized for GAPDH and β-actin levels in extracts of carotid plaques from 70 asymptomatic and 104 symptomatic patients.