Human TUFM ELISA Matched Antibody Pair

Eicosapentaenoic acid (EPA), an n-3 polyunsaturated fatty acid (PUFA), has been shown to decrease the risk of atherosclerosis by attenuating endothelial activation. In this study, we used mass spectrometry-based label-free quantitative proteomics to study the protective mechanisms of EPA and to identify key proteins that regulated by EPA in endothelial activation. Arachidonic acid (AA) was used as a control. HUVECs were pretreated with each of the two PUFAs, and then stimulated with TNF alpha as a model of endothelial activation. A total of 3391 proteins were identified, and 1958 proteins were quantified. Pearson's correlation coefficients revealed the excellent biological reproducibility of the proteomic results. Gene Ontology and KEGG enrichment analysis of differentially expressed proteins was performed, thus leading to the identification of the glutathione metabolism, oxidation reduction, and DNA replication as the most significantly enriched pathways. Seven key proteins were identified: elongation factor Tu (mitochondrial, TUFM), integrin alpha 6 (ITGA6), catalase (CAT), annexin A6 (ANXA6), heat shock 70 kDa protein 1A (HSPA1A), glutamate-cysteine ligase regulatory subunit (GCLM), and heme oxygenase 1 (HMOX1). Further connections among these proteins were also revealed by protein-protein interaction analysis. The mRNA levels of CAT, GCLM, and HMOX1 were verified with real-time PCR. The protein level of CAT was verified using Western blotting. This study is an in-depth proteomics analysis of EPA-treated cells and may provide possible insights into the molecular mechanisms of EPA's cytoprotective and atheroprotective effects. (C) 2017 Elsevier Inc. All rights reserved.

Profiles of sequence variants that influence gene transcription are very important for understanding mechanisms that affect phenotypic variation and disease susceptibility. Using genotypes at 1.4 million SNPs and a comprehensive transcriptional profile of 15,454 coding genes and 6,113 lincRNA genes obtained from peripheral blood cells of 298 Japanese individuals, we mapped expression quantitative trait loci (eQTLs). We identified 3,804 cis-eQTLs (within 500 kb from target genes) and 165 trans-eQTLs (>500 kb away or on different chromosomes). Cis-eQTLs were often located in transcribed or adjacent regions of genes; among these regions, 59 untranslated regions and 59 flanking regions had the largest effects. Epigenetic evidence for regulatory potential accumulated in public databases explained the magnitude of the effects of our eQTLs. Cis-eQTLs were often located near the respective target genes, if not within genes. Large effect sizes were observed with eQTLs near target genes, and effect sizes were obviously attenuated as the eQTL distance from the gene increased. Using a very stringent significance threshold, we identified 165 large-effect trans-eQTLs. We used our eQTL map to assess 8,069 disease-associated SNPs identified in 1,436 genome-wide association studies (GWAS). We identified genes that might be truly causative, but GWAS might have failed to identify for 148 out of the GWAS-identified SNPs; for example, TUFM (P = 3.3E-48) was identified for inflammatory bowel disease (early onset); ZFP90 (P = 4.4E-34) for ulcerative colitis; and IDUA (P = 2.2E-11) for Parkinson's disease. We identified four genes (P<2.0E-14) that might be related to three diseases and two hematological traits; each expression is regulated by trans-eQTLs on a different chromosome than the gene.