Genetic variation at chromosome 1p13 Genetic variation at chromosome 1p13.3 affects sortilin mRNA expression, cellular LDL- uptake and serum LDL levels which translates to the risk of coronary artery disease  Patrick Linsel-Nitschke, Jörg Heeren, Zouhair Aherrahrou, Petra Bruse, Christian Gieger, Thomas Illig, Holger Prokisch, Katharina Heim, Angela Doering, Annette Peters, Thomas Meitinger, H.-Erich Wichmann, Anke Hinney, Thomas Reinehr, Christian Roth, Jan. R. Ortlepp, Mouhidien Soufi, Alexander M. Sattler, Jürgen Schaefer, Klaus Stark, Christian Hengstenberg, Arne Schaefer, Stefan Schreiber, Florian Kronenberg, Nilesh J. Samani, Heribert Schunkert, Jeanette Erdmann  Atherosclerosis  Volume 208, Issue 1, Pages 183-189 (January 2010) DOI: 10.1016/j.atherosclerosis.2009.06.034 Copyright © 2009 Elsevier Ireland Ltd Terms and Conditions

Fig. 1 Association of SNP rs599839 with serum concentration of LDL-C. Presented are LDL-C levels stratified by rs599839 genotype in the population-based KORA F3 and S4 studies and in children and adolescents from the West German Obesity Cohort. The upper panel depicts each study population separately, whereas the lower panel shows LDL-C levels in different age groups. For the single-studies age and gender-adjusted means, standard errors and one-sided p-values from linear regression on LDL-C levels are displayed. For the pooled analysis of the population-based studies, KORA/MONICA F3, S4 and West German Obesity Cohort, mean, standard error and one-sided p-value from fixed effect linear regression model with adjustment for study, age and gender are displayed. Atherosclerosis 2010 208, 183-189DOI: (10.1016/j.atherosclerosis.2009.06.034) Copyright © 2009 Elsevier Ireland Ltd Terms and Conditions

Fig. 2 Forest-plot of rs599839 on CAD. Presented are odds ratios (OR) and confidence intervals (CI) per one G-allele under the assumption of an additive model. Boxes indicate the relative size of the samples. Single study analysis: one-sided asymptotic p-value of Cochran–Armitage trend test and corresponding OR and 90% CI. Pooled analysis: one-sided asymptotic p-value corresponding OR and 90% CI from logistic regression models with fixed effect adjustments for study. Atherosclerosis 2010 208, 183-189DOI: (10.1016/j.atherosclerosis.2009.06.034) Copyright © 2009 Elsevier Ireland Ltd Terms and Conditions

Fig. 3 Association of SNP rs599839 with expression levels of mRNAs for SORT1, CELSR2, PSRC1 and for LDL-R, RAP and LRP. Normalized mRNA-expression levels stratified by rs599839 genotype are displayed for three genes located at chromosome 1p13 in proximity to SNP rs599839 and for three genes closely related to LDL-metabolism. p-Values for the difference in mRNA expression between the AA compared to the GG genotype are from a two-tailed student's t-test. Atherosclerosis 2010 208, 183-189DOI: (10.1016/j.atherosclerosis.2009.06.034) Copyright © 2009 Elsevier Ireland Ltd Terms and Conditions

Fig. 4 (a) Overexpression of SORT1 in HEK-293 cells. The membrane was probed with the anti-human/mouse sortilin antibody detecting the sortilin transmembrane receptor by 100kDa. Two additional forms of sortilin were also detected around 80kDa. Four wells in the plates were transfected with each empty vector (pCMV6-XL5; mock) and vector carrying the cDNA from Sort 1 (pCMV6-XL5-Sort1) constructs, respectively. Two samples from each transfected well are represented (lanes SORT1 3–4, and mock 5–6). Lanes 1 and 2 correspond to Marker and Buffer, respectively. (b) Uptake of radiolabelled LDL- and RAP-particles into transfected HEK293-cells. Displayed are a dose response experiment for the uptake of 125I-LDL in transfected cells (upper left panel), a comparison of LDL and RAP uptake in transfected cells (upper right panel) and LDL and RAP uptake after unlabelled lipoprotein-lipase or RAP was added to the medium (lower left and right panel, respectively). Each experiment was performed at least in triplicate. Cell associated radioactivity per mg cell protein is displayed on the y-axis, type of DNA used for transfection and concentrations of radioactive LDL added as ligand into the medium are depicted on x-axis. For each experimental condition a two-tailed students t-test comparing mock- or SORT1-transfected cells are displayed. Atherosclerosis 2010 208, 183-189DOI: (10.1016/j.atherosclerosis.2009.06.034) Copyright © 2009 Elsevier Ireland Ltd Terms and Conditions

Fig. 5 Simplified hypothesis for the relationship between SNP rs599839, sortilin, LDL-levels and CAD risk. Atherosclerosis 2010 208, 183-189DOI: (10.1016/j.atherosclerosis.2009.06.034) Copyright © 2009 Elsevier Ireland Ltd Terms and Conditions