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

Similar documents
2.5. AMPK activity

Quantitative Real-Time PCR was performed as same as Materials and Methods.

G. Chinetti-Gbaguidi and B. Staels, UR 545 INSERM, Institut Pasteur de Lille and Université de Lille 2, Lille, France

Nature Medicine doi: /nm.3150

Materials and Methods Cell culture Foam cell formation Cellular uptake of DiI-OxLDL In vitro cholesterol efflux assays

Supplementary Figure S I: Effects of D4F on body weight and serum lipids in apoe -/- mice.

Table S1. Quantitative RT-PCR primers

High density lipoprotein metabolism

a. b. c. d. e. f. g. h. i. j. k. l. m. n. o. p.

Amniotic fluid stem cells provide considerable advantages in epidermal. regeneration: B7H4 creates a moderate inflammation

Potential Atheroprotective Effects of Ixmyelocel-T Cellular Therapy. Kelly J. Ledford, Nikki Murphy, Frank Zeigler, Ronnda L.

Peroxisome proliferator-activated receptors (PPARs) are

Figure 1. Effects of FGF21 on adipose tissue. (A) Representative histological. findings of epididymal adipose tissue (B) mrna expression of

Overexpression of STARD3 in human monocyte/macrophages induces an anti-atherogenic lipid phenotype

Gene expression was analyzed by real time PCR (SYBR GREEN PCR Master. Mix, Roche Applied Science) using specific oligonucleotides.

Cells and reagents. Synaptopodin knockdown (1) and dynamin knockdown (2)

Differential inhibition of macrophage foam-cell formation and atherosclerosis in mice by PPARα, β/δ, and γ

SUPPLEMENTARY INFORMATION

Plasma exposure levels from individual mice 4 hours post IP administration at the

Department of Pharmaceutical Sciences, School of Pharmacy, Northeastern University, Boston, MA 02115, USA 2

Commensal Bacteria at the Crossroad Between Cholesterol Homeostasis and Chronic Inflammation. in Atherosclerosis

Gene Polymorphisms and Carbohydrate Diets. James M. Ntambi Ph.D

Supplement Material. Spleen weight (mg) LN cells (X106) Acat1-/- Acat1-/- Mouse weight (g)

Atherosclerosis is a chronic inflammatory response to. Neopterin Counters Vascular Inflammation and Atherosclerosis ORIGINAL RESEARCH

IL-24 AND ITS ROLE IN WOUND HEALING

Increased CD36 protein as a response to defective insulin signaling in macrophages

Stewart et al. CD36 ligands promote sterile inflammation through assembly of a TLR 4 and 6 heterodimer

Lipid metabolism in familial hypercholesterolemia

Macrophages play essential roles in immunity and lipid. Regulation of Macrophage Functions by PPAR-, PPAR-, and LXRs in Mice and Men

Endothelial Dysfunction in Rheumatoid Arthritis: the Role of Monocyte Chemotactic

Supplementary Information. MicroRNA-33b knock-in mice for an intron of sterol regulatory

Original Article Regulation of macrophage cholesterol efflux and liver X receptor α activation by nicotine

Supplementary Figure 1. Repression of hepcidin expression in the liver of mice treated with

Summary and concluding remarks

Lipoproteins Metabolism Reference: Campbell Biochemistry and Lippincott s Biochemistry

Regulation of Lipid Homeostasis: Lipid Droplets

Intracellular MHC class II molecules promote TLR-triggered innate. immune responses by maintaining Btk activation

Supporting Information

JLR Papers In Press. Published on October 16, 2003 as Manuscript D JLR200

Ebrahim Abbasi Oshaghi 1,2

Autophagy involved in lipopolysaccharide-induced foam cell formation is mediated by adipose differentiation-related protein

Sphingolipid de novo synthesis: its relevance to metabolic diseases and cell polarity


Effect of High-fat or High-glucose Diet on Obesity and Visceral Adipose Tissue in Mice

Niacin Metabolism: Effects on Cholesterol

PPARg phosphorylation mediated by JNK MAPK: a potential role in macrophage-derived

1Why lipids cannot be transported in blood alone? 2How we transport Fatty acids and steroid hormones?

Supplementary Figure 1. PAQR3 knockdown inhibits SREBP-2 processing in CHO-7 cells CHO-7 cells were transfected with control sirna or a sirna

Leptin deficiency suppresses progression of atherosclerosis in apoe-deficient mice

Supplemental Table I.

SUPPLEMENTARY INFORMATION

Serafino et al. Thymosin α1 activates complement receptor-mediated phagocytosis in human monocyte-derived macrophages. SUPPLEMENTARY FIGURES

Aspergillus fumigatus activates PAR-2 and skews toward a Th2 bias in airway epithelial cells.

Review. Lipid droplet associated proteins: an emerging role in atherogenesis. Histology and Histopathology

EFFECTS OF PPARγ LIGANDS ON ATHEROSCLEROSIS AND CARDIOVASCULAR DISEASE

The uptake of lipids by macrophages (M ) resulting in

Human and mouse T cell regulation mediated by soluble CD52 interaction with Siglec-10. Esther Bandala-Sanchez, Yuxia Zhang, Simone Reinwald,

Gallic acid prevents isoproterenol-induced cardiac hypertrophy and fibrosis through regulation of JNK2 signaling and Smad3 binding activity

Lipid/Lipoprotein Structure and Metabolism (Overview)

Supplementary Figure 1: TSLP receptor skin expression in dcssc. A: Healthy control (HC) skin with TSLP receptor expression in brown (10x


BIOFENOLI ED INIBIZIONE DELL UPTAKE DI oxldl DA PARTE DI CELLULE MACROFAGICHE UMANE E MURINE

Supplementary Information

ZL ZDF ZDF + E2 *** Visceral (g) ZDF

Cell isolation. Spleen and lymph nodes (axillary, inguinal) were removed from mice

Protection against doxorubicin-induced myocardial dysfunction in mice by cardiac-specific expression of carboxyl terminus of hsp70-interacting protein

Supplementary Table 1. List of primers used in this study

Supplementary Figure 1. Supernatants electrophoresis from CD14+ and dendritic cells. Supernatants were resolved by SDS-PAGE and stained with

Supplementary Figure 1.

Dietary α-linolenic acid-rich flaxseed oil prevents against alcoholic hepatic steatosis

Nature Immunology: doi: /ni Supplementary Figure 1

Abstract: I. A ims Aim 1:

Supplemental information

Supplemental Figure 1. Western blot analysis indicated that MIF was detected in the fractions of

Supplementary Fig. 1. GPRC5A post-transcriptionally down-regulates EGFR expression. (a) Plot of the changes in steady state mrna levels versus

SUPPLEMENTARY INFORMATION

Role of CD36, the Macrophage Class B Scavenger Receptor, in Atherosclerosis

Therapeutic effect of baicalin on experimental autoimmune encephalomyelitis. is mediated by SOCS3 regulatory pathway

Supplementary Table 2. Conserved regulatory elements in the promoters of CD36.

Vadim Ivanov, M.D., Ph.D. Micronutrients in controlling INFLAMMATION Webinar June 12, 2012

SR-BI inhibits ABCG1-stimulated net cholesterol efflux from cells to plasma HDL

Microglia, Inflammation, and FTD

Chapter VIII: Dr. Sameh Sarray Hlaoui

Supplementary Data. (continued)

Supplementary methods:

Doctoral Degree Program in Marine Biotechnology, College of Marine Sciences, Doctoral Degree Program in Marine Biotechnology, Academia Sinica, Taipei,

Supplementary Figure 1.

Supplemental Material. Results

Hmgcoar AGCTTGCCCGAATTGTATGTG TCTGTTGTAACCATGTGACTTC. Cyp7α GGGATTGCTGTGGTAGTGAGC GGTATGGAATCAACCCGTTGTC

Anti-atherogenic actions of dihomo-gamma-linolenic acid in macrophages

Writing Effective Grant Proposals

Supplementary table I. Real-time primers used in the study. The fold change was obtained by

Supplementary Materials for

Effects of Gelsolin on Macrophage Inflammatory Responses to Implant Wear Debris

Supplementary Figure 1. Deletion of Smad3 prevents B16F10 melanoma invasion and metastasis in a mouse s.c. tumor model.

Atherosclerosis Compendium

Supporting Information. Supporting Tables. S-Table 1 Primer pairs for RT-PCR. Product size. Gene Primer pairs

Spherical Nucleic Acids For Advanced Wound Healing Applications Chad A. Mirkin

Programmed necrosis, not apoptosis, is a key mediator of cell loss and DAMP-mediated inflammation in dsrna-induced retinal degeneration

Supplementary information. The proton-sensing G protein-coupled receptor T-cell death-associated gene 8

Transcription:

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

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. 2002. 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 32:2979-87. 2. Draude, G., and R. L. Lorenz. 2000. TGF-beta1 downregulates CD36 and scavenger receptor A but upregulates LOX-1 in human macrophages. American Journal of Physiology - Heart & Circulatory Physiology 278:H1042-8. 3. Eguchi, A., A. Murakami, and H. Ohigashi. 2006. Nobiletin, a citrus flavonoid, suppresses phorbol ester-induced expression of multiple scavenger receptor genes in THP-1 human monocytic cells. FEBS Letters 580:3321-8. 4. Singh, N. N., and D. P. Ramji. 2006. 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 26:1323-9. 5. Kaplan, R., X. Gan, J. G. Menke, S. D. Wright, and T. Q. Cai. 2002. Bacterial lipopolysaccharide induces expression of ABCA1 but not ABCG1 via an LXR-independent pathway. Journal of Lipid Research 43:952-9. 6. Chinetti-Gbaguidi, G., E. Rigamonti, L. Helin, A. L. Mutka, M. Lepore, J. C. Fruchart, V. Clavey, E. Ikonen, S. Lestavel, and B. Staels. 2005. Peroxisome proliferator-activated receptor alpha controls cellular cholesterol trafficking in macrophages. Journal of Lipid Research 46:2717-25. 7. Chinetti, G., S. Lestavel, J. C. Fruchart, V. Clavey, and B. Staels. 2003. Peroxisome proliferator-activated receptor alpha reduces cholesterol esterification in macrophages. Circulation Research 92:212-7. 8. Larigauderie, G., C. Cuaz-Perolin, A. B. Younes, C. Furman, C. Lasselin, C. Copin, M. Jaye, J. C. Fruchart, and M. Rouis. 2006. Adipophilin increases triglyceride storage in human macrophages by stimulation of biosynthesis and inhibition of beta-oxidation. FEBS Journal 273:3498-510. 2

9. Yamanishi, Y., D. L. Boyle, M. Clark, R. A. Maki, M. D. Tortorella, E. C. Arner, and G. S. Firestein. 2002. Expression and regulation of aggrecanase in arthritis: the role of TGF-beta. Journal of Immunology 168:1405-12. 10. 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. 2009. TNF-alpha stimulates the ACAT1 expression in differentiating monocytes to promote the CE-laden cell formation. Journal of Lipid Research 50:1057-67. 11. Hickman, S. E., E. K. Allison, and J. El Khoury. 2008. Microglial dysfunction and defective beta-amyloid clearance pathways in aging Alzheimer's disease mice. Journal of Neuroscience 28:8354-60. 12. Gafencu, A. V., M. R. Robciuc, E. Fuior, V. I. Zannis, D. Kardassis, and M. Simionescu. 2007. Inflammatory signaling pathways regulating ApoE gene expression in macrophages. Journal of Biological Chemistry 282:21776-85. 13. Harvey, E. J., N. Li, and D. P. Ramji. 2007. 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 27:806-12. 3

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<0.001. 4

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

A Supplement Figure II 120 % DiI-AcLDL uptake 100 80 60 40 20 ** ** *** *** 0 B No DiI- AcLDL Untreated (1 ng/ml) (5 ng/ml) (10 ng/ml) (20 ng/ml) 120 % DiI-AcLDL uptake 100 80 60 40 20 * * ** *** 0 No DiI- AcLDL Untreated 1hr 3hr 6hr 24hr