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1 SUPPLEMENTL MTERIL Supplemental Figure I body weight [g] fasting blood glucose [mg/dl] C total cholesterol [mg/dl] WT Has -/- Supplemental Figure I: Has-deficient mice exhibited no apparent developmental abnormalities and had normal life spans., ody weight of adult mice, n =, 5., 6h fasting blood glucose levels; n = 8, 6, C, total cholesterol; n = 6,. Data are means ± SEM.
2 Supplemental Figure II WT, ligated vs. non-ligated 86 differentially expressed transcripts p (corr) <.5 Has -/-, ligated vs. non-ligated 65 differentially expressed transcripts p (corr) <.5 Supplemental Figure II: Comparison of carotid gene expression in ligated versus nonligated carotid arteries reveals more differentially expressed genes in WT mice. Five days after surgery ligated carotid arteries and non-ligated control carotid arteries were harvested from Has-deficient mice and WT controls. Gene expression profiles were assessed using whole genome microarrays. Depiction of the number of differentially expressed genes with overlapping and distinct regulation in Has -/- and WT mice. Differential gene expression was statistically determined by moderated t tests, corrected for multiple testing (enjamini-hochberg FDR); n =,4, p (corr) <.5.
3 Supplemental Figure III non-ligated ligated non-ligated WT ligated - Has -/- C 4 D 4 WT Has -/- -log corrected p value -log corrected p value log [fold change] log [fold change] Supplemental Figure III: Hierarchical clustering and volcano plots show a different pattern of differentially regulated genes after carotid artery ligation comparing Has-deficient and WT mice. Five days after surgery ligated carotid arteries and non-ligated control carotid arteries were harvested from Has-deficient mice and WT controls. Changes in carotid gene expression were analyzed by microarrays comparing ligated carotid arteries with non-ligated controls.,, Hierarchical clustering analyses for WT () (86 transcripts) and Has-deficient () (65 transcripts) animals show grouping of differentially expressed transcripts of the non-ligated and ligated samples. C, D, Volcano plots showing fold changes and significances for the detected genes in WT () and Has-deficient () mice. Significantly (p (corr) <.5) regulated genes are shown in red. Differential gene expression was statistically determined by moderated t tests, corrected for multiple testing (enjamini-hochberg FDR), n =,4, p (corr) <.5.
4 Supplemental Figure IV migration [fold of pcl] * [ H]-thymidine-incorporation [fold of pcl].5..5 * pcl HSoe PDGF- [ ng/ml] PDGF- [ ng/ml] Supplemental Figure IV: Overexpression of HS increases migration and proliferation in VSMC. Overexpression of HS (HSoe) induced migration,, and DN synthesis,, as determined by [ H]-thymidine incorporation in VSMC. n = -6, * p <.5 vs. control.
5 Supplemental Table I Pathway Hypertrophic cardiomyopathy (HCM) Regulation of actin cytoskeleton Dilated cardiomyopathy Focal adhesion Hematopoietic cell lineage ECM-receptor interaction rrhythmogenic right ventricular cardiomyopathy (RVC) xon guidance Vascular smooth muscle contraction Pathways in cancer Phosphatidylinositol signaling system Calcium signaling pathway dherens junction MPK signaling pathway Neurotrophin signaling pathway Gap junction Viral myocarditis Endocytosis Long-term potentiation (Synapses) Prion diseases Progesterone-mediated oocyte maturation Inositol phosphate metabolism Enrichment p-value 4,44E-9,E-7,E-7,7E-7,7E-7 8,67E-7,69E-5 7,4E-5,94E-4,5E-,E- 4,8E- 9,E-,6E-,5E-,46E-,77E-,96E-,E-,8E-,65E-,7E- Supplemental Table I: KEGG pathways exclusively regulated in carotid arteries of WT mice. Total RN was isolated from carotid arteries and subjected to whole genome microarray analysis. Indicated are only those transcriptional pathways that were regulated in WT, but not in Has- deficient animals.
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