Supplemental Materials

Supplementary Figure 1: Endothelial cell sample characterization

A. Nuclear staining en face  with Hoechst 33258. EC displayed alignment and morphology characteristic of their local hemodynamic environment. B. H & E stained histology sections. Histology was consistent with that of normal vessels and there was no evidence of pathology associated with either flow region.  Staining for oil red-O (not shown) was negative. C. Staining with EC-specific PECAM-1 (green) and SMC-specific a -actin (red) antibodies. Pure endothelial populations (>99%) were rapidly and routinely isolated.

Table II (Online Version): Differentially Expressed Genes by Region within Animal Group

DF-A = disturbed flow region (aortic arch), UF-A = undisturbed flow region (descending thoracic aorta), UF-C = undisturbed flow region (common carotid), ALL = across all animals, HCM = high cholesterol males, NM = normal males, NF = normal females, FDR = false discovery rate

Table III (Online Version): Differentially Expressed Genes by Risk Factor (Gender or Diet) within Region

DF-A = disturbed flow region (aortic arch), UF-A = undisturbed flow region (descending thoracic aorta), UF-C = undisturbed flow region (common carotid), HCM = high cholesterol males, NM = normal males, NF = normal females, FDR = false discovery rate

Supplementary Figure 2: Venn diagram showing the distribution of differentially expressed genes (DF-A vs UF-A) identified by PaGE analysis (FDR = 15%, confidence ≥ 85% within each animal subgroup (n=5)

HCM = high cholesterol males, NM = normal males, NF = normal females

Table IV (Online Version): Differentially expressed genes (DF-A vs UF-A) by biological classification

DF-A = disturbed flow region (aortic arch), UF-A = undisturbed flow region (descending thoracic aorta), ALL = across all animals, HCM = high cholesterol males, NM = normal males, NF = normal females, FDR = false discovery rate

Supplementary Table I: QRT-PCR Results

* indicates significantly different from 1.00

HCM = high cholesterol males, NM = normal males, NF = normal females

Supplementary Table II: Over-represented biological categories identified by EASE analysis

DF-A = disturbed flow region (aortic arch), UF-A = undisturbed flow region (descending thoracic aorta), ALL = across all animals, HCM = high cholesterol males, NM = normal males, NF = normal females

Supplementary Table III: Identification of Biological Themes using Ingenuity's Pathways Analysis

DF-A = disturbed flow region (aortic arch), UF-A = undisturbed flow region (descending thoracic aorta), ALL = across all animals, HCM = high cholesterol males, NM = normal males, NF = normal females