Adrenalectomy does not alter the size (A) or collagen content (B) of atherosclerotic lesions in APOE knockout mice.

<p>Representative neutral lipid (Oil red O) and collagen (Trichrome: blue) stainings of early (C) and advanced (D) aortic root lesions. Separate dots in figures A and B represent values for each individual mouse, while horizontal lines indicate the respective group averages.</p

Adrenalectomy in APOE knockout mice does not alter the absolute thymus weight, but increases the thymic CD4/CD8 double positive lymphocyte fraction.

<p>Values represent means+SEM of 5 (FACS data) or 10-15 (thymus weight) mice per group. * P<0.05.</p

Plasma postprandial triglyceride response (A), cholesterol levels (B), and lipoprotein profiles (C) in adrenalectomized (ADX) and SHAM-operated APOE knockout mice.

<p>VLDL, very low-density lipoprotein; LDL, low-denisty lipoprotein; HDL, high-density lipoprotein. Values represent means+SEM or 5 mice per group. * P<0.05, ** P<0.01, *** P<0.001.</p

<i>In vitro</i> cellular cholesterol efflux.

<p>Cholesterol efflux from bone marrow-derived macrophages (BMDM) to BSA (A), apoA-I (10 µg/mL; B) and HDL (50 µg/mL; C) was determined after 24 h. Data are expressed as the percentage of radioactivity released into the medium. Values represent the means of n≥3 mice ± SEM per group. ***p<0.001 as compared to WT transplanted controls.</p

Successful disruption of macrophage ABCA1 and apoE by bone marrow transplantation.

<p>RT-PCR on genomic DNA of bone marrow isolated from ABCA1 and/or apoE KO transplanted LDLr KO mice and their WT transplanted controls at 17 weeks after transplantation.</p

Hepatic gene expression levels and circulating IL-6 levels.

<p>mRNA expression of TNFα (A), and IL-6 (B) in liver from bone marrow transplanted LDLr KO mice. Gene expression analysis was performed by RT-PCR. Values are relative to the average expression of housekeeping genes and expressed as fold induction compared to WT transplanted controls. Values are mean ± SEM, n≥3 mice per group as compared to WT. Increased circulating IL-6 levels in LDLr KO mice transplanted with ABCA1/apoE double KO bone marrow (C). Serum from the transplanted LDLr KO mice was collected at 17 weeks after transplantation including 9 weeks of WTD feeding. Values are mean ± SEM, n≥8 mice per group. *p<0.05, **p<0.01. ***p<0.001 as compared to WT transplanted controls. ^#p<0.05 as compared to ABCA1 KO transplanted animals.</p

Analysis of atherosclerosis in the aortic arch of bone marrow transplanted LDLr KO mice after 9 weeks of HF/HC diet feeding.

<p>Relative plaque area in the aortic arch was quantified <i>en face</i> calculating the ratio of atherosclerotic lesions to the surface of the entire aortic arch (left panel). Representative pictures (right panels). **P<0.01 as compared to WT transplanted controls.</p

Lipoprotein profiles from ABCA1 and/or apoE KO transplanted LDLr KO mice and their WT transplanted controls.

<p>Fifty µL samples of serum, pooled from 2 mice per genotype, collected after 9 weeks of HF/HC feeding, were subjected to FPLC fractionation. Distribution of serum FC and TC over the fractions (25 µL) was determined enzymatically. Values represent the means of n≥5 samples per group. 1 sample corresponds to pooled serum of 2 mice.</p

Analysis of atherosclerosis in the aortic root of bone marrow transplanted LDLr KO mice after 9 weeks of HF/HC diet feeding.

<p>Quantification of atherosclerotic lesion sizes in the aortic root and TC levels in bone marrow transplanted LDLr KO mice after 9 weeks of HF/HC diet feeding (A). Lesion size divided by TC levels show cholesterol independent effects on lesion development (B). Representative cross-sections, stained with oil red-O (magnification 50×; lower panels).Values are the means of 10 aortic root sections of individual mice (n≥11 mice ± SEM per group. **p<0.01, ***p<0.001 as compared to WT transplanted controls, ^#p<0.05 as compared to ABCA1 KO transplanted animals.</p

ApoE protein content in VLDL and LDL fractions from ABCA1 and/or apoE KO transplanted LDLr KO mice and their WT transplanted controls.

<p>Western blots showing apoE protein expression in VLDL (A) and LDL (B) isolated from ABCA1 KO and/or apoE KO transplanted LDLr KO animals and WT transplanted controls after feeding the HF/HC diet for 9 weeks. Samples were adjusted for cholesterol content. The density of the bands was determined using quantification software (Image J). n≥5 samples per group (pooled). WT values were set at 100%.</p