Supplementary Table I - Primers used for real-time quantitative PCR and RT-PCR

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1 Supplement Supplementary Table I - Primers used for real-time quantitative PCR and RT-PCR Gene Forward Primer (5-3 ) Reverse primer (5-3 ) Reference Human ST2 CTTGATTGATAAACAGAATG CTGATCCAGATACTGTTGAA [1] Human SR-A CCAGGGACATGGAATGCAA CCAGTGGGACCTCGATCTCC [2] Human CD36 GAGAACTGTTATGGGGCTAT TTCAACTGGAGAGGCAAAGG [2] Human SR-BI TGATGATGGAGAATAAGCCCAT TGACCGGGTGGATGTCCAGGAAC [3] Human ApoE TTCCTGGCAGGATGCCAGGC GGTCAGTTGTTCCTCCAGTTC [4] Human ABCA-1 TGTCCAGTCCAGTAATGGTTCTGT AAGCGAGATATGGTCCGGATT [5] Human ABCG-1 TGCAATCTTGTGCCATATTTGA CCAGCCGACTGTTCTGATCA [6] Human NPC-1 CTTAGTGCAGGAACTCTGTCCAGG TCCACATCACGGCAGGCATTGTAC [6] Human NPC-2 GGTTTGTCTTGTGACCGC AGGAATGTAGCTGCCAGG [6] Human CPT-1 ACAGTCGGTGAGGCCTCTTATGAA TCTTGCTGCCTGAATGTGAGTTGG [7] Human ADRP TGTGAGATGGCAGAGAACGGT CTGCTCACGAGCTGCATCATC [8] Human ACAT-1 GATGAAGGAAGGCTGGTGC GGAAGCTGGTGGCAGTGTAT [9] Human NCEH CACTCCTGCTGACTTGACCA CATCCCCTGTGCTGAAGAAT - Human GAPDH GAAGGTGAAGGTCGGAGTC GAAGATGGTGATGGGATTTC [10] Mouse SR-A TGAACGAGAGGATGCTGACTG GGAGGGGCCATTTTTAGTGC [11] Mouse CD36 GAACCACTGCTTTCAAAAACTGG TGCTGTTCTTTGCCACGTCA [11] Mouse SR-BI TTTGGAGTGGTAGTAAAAAGGGC TGACATCAGGGACTCAGAGTAG [11] Mouse ApoE ACAGATCAGCTCGAGTGGCAAA ATCTTGCGCAGGTGTGTGGAGA [12] Mouse ABCA-1 AGTGATAATCAAAGTCAAAGGCACAC AGCAACTTGGCACTAGTAACTCTG [5] Mouse ABCG-1 TTCATCGTCCTGGGCATCTT CGGATTTTGTATCTGAGGACGAA [5] Mouse β-actin TGGAGAAGAGCTATGAGCTGCCTG GTGCCACCAGACAGCACTGTGTTG [13] 1

2 References 1. Lecart, S., N. Lecointe, A. Subramaniam, S. Alkan, D. Ni, R. Chen, V. Boulay, J. Pene, K. Kuroiwa, S. Tominaga, and H. Yssel Activated, but not resting human Th2 cells, in contrast to Th1 and T regulatory cells, produce soluble ST2 and express low levels of ST2L at the cell surface. European Journal of Immunology 32: Draude, G., and R. L. Lorenz TGF-beta1 downregulates CD36 and scavenger receptor A but upregulates LOX-1 in human macrophages. American Journal of Physiology - Heart & Circulatory Physiology 278:H Eguchi, A., A. Murakami, and H. Ohigashi Nobiletin, a citrus flavonoid, suppresses phorbol ester-induced expression of multiple scavenger receptor genes in THP-1 human monocytic cells. FEBS Letters 580: Singh, N. N., and D. P. Ramji Transforming growth factor-beta-induced expression of the apolipoprotein E gene requires c-jun N-terminal kinase, p38 kinase, and casein kinase 2. Arteriosclerosis, Thrombosis & Vascular Biology 26: Kaplan, R., X. Gan, J. G. Menke, S. D. Wright, and T. Q. Cai Bacterial lipopolysaccharide induces expression of ABCA1 but not ABCG1 via an LXR-independent pathway. Journal of Lipid Research 43: Chinetti-Gbaguidi, G., E. Rigamonti, L. Helin, A. L. Mutka, M. Lepore, J. C. Fruchart, V. Clavey, E. Ikonen, S. Lestavel, and B. Staels Peroxisome proliferator-activated receptor alpha controls cellular cholesterol trafficking in macrophages. Journal of Lipid Research 46: Chinetti, G., S. Lestavel, J. C. Fruchart, V. Clavey, and B. Staels Peroxisome proliferator-activated receptor alpha reduces cholesterol esterification in macrophages. Circulation Research 92: Larigauderie, G., C. Cuaz-Perolin, A. B. Younes, C. Furman, C. Lasselin, C. Copin, M. Jaye, J. C. Fruchart, and M. Rouis Adipophilin increases triglyceride storage in human macrophages by stimulation of biosynthesis and inhibition of beta-oxidation. FEBS Journal 273:

3 9. Yamanishi, Y., D. L. Boyle, M. Clark, R. A. Maki, M. D. Tortorella, E. C. Arner, and G. S. Firestein Expression and regulation of aggrecanase in arthritis: the role of TGF-beta. Journal of Immunology 168: Lei, L., Y. Xiong, J. Chen, J. B. Yang, Y. Wang, X. Y. Yang, C. C. Chang, B. L. Song, T. Y. Chang, and B. L. Li TNF-alpha stimulates the ACAT1 expression in differentiating monocytes to promote the CE-laden cell formation. Journal of Lipid Research 50: Hickman, S. E., E. K. Allison, and J. El Khoury Microglial dysfunction and defective beta-amyloid clearance pathways in aging Alzheimer's disease mice. Journal of Neuroscience 28: Gafencu, A. V., M. R. Robciuc, E. Fuior, V. I. Zannis, D. Kardassis, and M. Simionescu Inflammatory signaling pathways regulating ApoE gene expression in macrophages. Journal of Biological Chemistry 282: Harvey, E. J., N. Li, and D. P. Ramji Critical role for casein kinase 2 and phosphoinositide-3-kinase in the interferon-gamma-induced expression of monocyte chemoattractant protein-1 and other key genes implicated in atherosclerosis. Arteriosclerosis, Thrombosis & Vascular Biology 27:

4 Supplementary Figure Legends Supplement Figure I ST2 mrna expression in THP-1 macrophages and HMDMs ST2 RT-PCR was performed on cdna generated from THP-1 macrophages and HMDMs using primers detailed in supplementary table I. GAPDH was used as an internal control (20 cycles, 60 C annealing temperature). The PCR products were size fractionated by agarose gel electrophoresis and analyzed using a Syngene gel documentation system (GRI). RT indicates where reverse transcriptase was omitted in the cdna synthesis step. M indicates molecular weight markers. Data is indicative of three separate experiments. Supplement Figure II reduces AcLDL uptake by THP-1 macrophages in a time and concentration-dependent manner DiI-AcLDL uptake was measured in 24 hour 160nM PMA-differentiated THP-1 macrophages incubated with a range of concentrations (1-20 ng/ml) of for 24 hours (n=3) (A) or over a range of time points (1-24 hours) with (10 ng/ml) (n=3) (B). Data represents mean+sd. Student s t- test, * P<0.05; ** P<0.01; *** P<

5 Supplement Figure I THP-1 HMDMs -RT ST2 GAPDH 4

6 A Supplement Figure II 120 % DiI-AcLDL uptake ** ** *** *** 0 B No DiI- AcLDL Untreated (1 ng/ml) (5 ng/ml) (10 ng/ml) (20 ng/ml) 120 % DiI-AcLDL uptake * * ** *** 0 No DiI- AcLDL Untreated 1hr 3hr 6hr 24hr

2.5. AMPK activity

2.5. AMPK activity Supplement Fig. A 3 B phos-ampk 2.5 * Control AICAR AMPK AMPK activity (Absorbance at 45 nm) 2.5.5 Control AICAR Supplement Fig. Effects of AICAR on AMPK activation in macrophages. J774. macrophages were