EMT and ECM. Jesse Liang, Ph.D. Sample & Assay Technologies

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1 EMT and ECM Jesse Liang, Ph.D. 1

2 Epithelial Cells.1. Closely adjoined.2. Polarized.Epithelial Markers: E-Cadherin (adherens junctions) Claudins (tight junctions) Occludin (tight junctions) Desmoplakin (desmosomes) Cytokeratin-8, -18 and -19 Mucin-1 There are 5 different types of cell junctions. They are tight junctions, adherens junctions, desmosomes, hemidesmosomes, and gap junctions

3 Mesenchymal Cells.1. Not adjoined.2. No polarity.mesenchymal Markers:.Vimentin.N-Cadherin.Fibronectin.Vitronectin.FSP1(fibroblast-specific protein 1).Smooth-muscle actin.fgfr2 IIIb and IIIc splice variants - 3 -

4 EMT (Epithelial-Mesenchymal Transition).Epithelial cells can convert into mesenchymal cells by a process known as EMT, which disrupts cell-cell adhesion and cell-ecm adhesion..* Embryogenesis and development.* Cancer.* Fibrosis - 4 -

5 The Nature of EMT Transient (reversible) Highly context-specific A vicious cycle in pathological conditions - 5 -

Activators and Suppressors of EMT Epithelial-Mesenchymal Transitions in Development and Disease Cell (2009) 139, 871-890.

6 Activators and Suppressors of EMT Epithelial-Mesenchymal Transitions in Development and Disease Cell (2009) 139, highly context-specific TGFβ, FGF, EGF families, HGF; Src, GTPase family Ras, Rho, Rac; Snail, Slug, Twist, ZEB, NFκB Extracellular Cytoplasmic Nuclear - 6 -

7 EMT Signaling A central target is E-cadherin Wilm s tumor gene 1, In heart development Oxygen-dependent gene expression in development and cancer: lessons learned from the Wilm s tumor gene, WT1. Front Mol Neurosci (2011), 4:

8 EMT and Cancer Progression of most carcinomas is associated with the acquisition of mesenchymal phenotype. Cells with an EMT phenotype induced by different factors are rich sources for cancer stem-like cells. Moreover, induction of EMT in tumor cells not only promotes invasion and metastasis but also contributes to drug resistance

9 Induction of EMT Generates Stem-Like Cells Breast cancer initiating cells are CD44 + CD24 - cells. EMT phenotype EMT induction Mani SA, et al. The Epithelial-Mesenchymal Transition Generates Cells with Properties of Stem Cells. Cell (2008), 133, Leukemia initiating cells are CD34+CD38 cells. Colon cancer initiating cells are CD133+ cells. Brain cancer initiating cells are CD133+ cells. Prostate cancer initiating cells are CD44+α2β1+ cells

10 mirnas Link EMT to Stem-Like Cells in Human Cancers mir-200 family and ZEB1/2 mir-200a * Knockdown of Akt-1 decreases the expression of mir-200 family including mir- 200a, and increases mammosphere forming ability in breast cancer mir-200b * mir-200b inhibits expression of ZEB1, ZEB2, Lin28B and Notch1 in prostate cancer * mir-200b targets Suz12 and contributes to cancer stem cells maintenance in breast cancer mir-200c * mir-200c inhibits expression of ZEB1, ZEB2 and Bmi1 in breast cancer; * mir-200c inhibits expression of ZEB1, Sox2, Bmi1 and KLF4 in pancreatic cancer mir-183 * ZEB1 represses mir-183 expression, which increases the expression of Bmi1 and KLF4 in pancreatic cancer mir-203 * ZEB1 represses mir-203 expression, which increases the expression of Bmi1 and KLF4 in pancreatic cancer

11 EMT and Fibrosis Fibrosis is characterized by the presence of an excess of fibrous connective tissue in an organ, and in particular by an excessive deposition of collagen I. Renal fibrosis, for example, has been associated with the activation of interstitial fibroblasts, which give rise to collagen secreting myofibroblasts. In addition, myofibroblasts can also originate from renal tubular epithelial and endothelial cells that undergo EMT. Activation of Snail1, a well known EMT inducer, leads to renal fibrosis and renal failure in animal models. High Snail1 expression and evidence of EMT has also been found in the kidneys of patients with renal fibrosis (Boutet et al, 2006)

12 Cancer and Fibrosis are (Induced by) Inflammation In the context of a chronic inflammatory condition, TGFβ1 and hypoxia activate EMT that converges in the activation of NFκB, which is also induced by the inflammatory cytokines and oxidative stress

13 Inflammation, Oxidative Stress and Hypoxia in EMT Induction Inflammation and EMT: an alliance towards organ fibrosis and cancer progression. EMBO Molecular Medicine (2009), 1,

Contamination Control Reverse Transcription

14 PCR Array Introduction 84 Pathway-Specific Genes of Interest 5 Housekeeping Genes Genomic DNA Contamination Control Reverse Transcription Controls (RTC) n=3 Positive PCR Controls (PPC) n=3-14 -

15 EMT PCR Arrays Genes Up-Regulated During EMT: AHNAK, BMP1, CALD1, CAMK2N1, CDH2, COL1A2, COL3A1, COL5A2, FN1, FOXC2, GNG11, GSC, IGFBP4, ITGA5, ITGAV, MMP2, MMP3, MMP9, MSN, SERPINE1, SNAI1, SNAI2, SNAI3, SOX10, SPARC, STEAP1, TCF4, TIMP1, TMEFF1, TMEM132A, TWIST1, VCAN, VIM, VPS13A, WNT5A, WNT5B. Genes Down-Regulated During EMT: CAV2, CDH1 (E-cadherin), DSP, FGFBP1, IL1RN, KRT19, MST1R, NUDT13, OCLN, PPPDE2, RGS2, SPP1 (Osteopontin), TFPI2, TSPAN13. Differentiation & Development: AKT1, BMP1, BMP2, BMP7, COL3A1, COL5A2, CTNNB1, DSP, ERBB3, F11R, FOXC2, FZD7, GSC, JAG1, KRT14, MST1R, NODAL, NOTCH1, PTP4A1, SMAD2, SNAI1, SNAI2, SOX10, TGFB2, TGFB3, TMEFF1, TWIST1, VCAN, WNT11, WNT5A, WNT5B. Morphogenesis: CTNNB1, FOXC2, JAG1, RAC1, SMAD2, SNAI1, SOX10, TGFB1, TGFB2, TGFB3, TWIST1, WNT11, WNT5A. Cell Growth & Proliferation: AKT1, BMP1, BMP7, CAV2, CTNNB1, EGFR, ERBB3, FGFBP1, FOXC2, IGFBP4, ILK, JAG1, MST1R, NODAL, PDGFRB, TGFB1, TGFB2, TGFB3, TIMP1, VCAN, ZEB1. Migration & Motility: CALD1, CAV2, EGFR, FN1, ITGB1, JAG1, MSN, MST1R, NODAL, PDGFRB, RAC1, STAT3, TGFB1, VIM. Cytoskeleton: CAV2, KRT7, MAP1B, PLEK2, RAC1, VIM. Extracellular Matrix & Cell Adhesion: BMP1, BMP7, CDH1 (E-cadherin), CDH2 (N-cadherin), COL1A2, COL3A1, COL5A2, CTNNB1, DSC2, EGFR, ERBB3, F11R, FN1, FOXC2, ILK, ITGA5, ITGAV, ITGB1, MMP2, MMP3, MMP9, PTK2, RAC1, SERPINE1 (PAI-1), SPP1 (Osteopontin), TGFB1, TGFB2, TIMP1, VCAN. Signaling Pathways: Estrogen Receptor: CAV2, ESR1 (ERa), KRT19, TGFB3. G-Protein Coupled Receptor: AKT1, FZD7, GNG11, RAC1, RGS2. Integrin-Mediated: COL3A1, ILK, ITGA5, ITGAV, ITGB1, PTK2. Notch: FOXC2, JAG1, NOTCH1. Receptor Tyrosine Kinase: EGFR, ERBB3, PDGFRB, RGS2, SPARC. TGFß / BMP: BMP1, BMP2, BMP7, COL3A1, SMAD2, TGFB1, TGFB2, TGFB3. WNT: CTNNB1, FZD7, GSK3B, WNT11, WNT5A, WNT5B. Transcription Factors: CTNNB1, ESR1 (ERa), FOXC2, GSC, NOTCH1, SIP1, SMAD2, SNAI2, SNAI3, SOX10, STAT3, TCF3, TCF4, TWIST1, ZEB1, ZEB

16 EMT Methylation PCR Arrays Genes Down-Regulated During EMT: CDH1, DSP, KRT19, MST1R, OCLN, PPPDE2, RGS2, TSPAN13. Differentiation & Development: MST1R, PTP4A1, SMAD4. Cell Growth & Proliferation: GAB1, MST1R, SEH1L, SMAD4. Extracellular Matrix & Cell Adhesion: CDH1, CTNNAL1, DSC2, EPCAM, NID2. Signal Transduction: GAB1, KRT19, MAP3K5, RGS2, SMAD4, TGIF1. Cytoskeleton: CTNNAL1, KRT7, PLEK2. Other Genes: PLSCR1, YES

17 EMT ChIP PCR Arrays Profile Histone Codes Genes Up-Regulated During EMT: AHNAK, BMP1, CALD1, CDH2 (N-cadherin), COL1A2, COL3A1, COL5A2, FN1, FOXC2, GNG11, GSC, IGFBP4, ITGA5, ITGAV, MMP2 (Gelatinase A), MMP3, MMP9 (Gelatinase B), MSN, SERPINE1 (PAI-1), SNAI1, SNAI2, SNAI3, SOX10, SPARC, STEAP1, TCF4, TIMP1, TMEFF1, TMEM132A, TWIST1, VCAN, VIM, VPS13A, WNT5A, WNT5B. Genes Down-Regulated During EMT: CAV2, CDH1 (E-cadherin), DSP, FGFBP1, IL1RN, KRT19, MITF, MST1R, NUDT13, PPPDE2, RGS2, SPP1 (Osteopontin), TFPI2, TSPAN13. Genes with Known Histone Modifications during EMT: Increased H3K4me3: AHNAK, AKT1, BMP1, CALD1, CAV2, CDH2 (N-cadherin), CTNNB1, FN1, FZD7, GNG11, GSK3B, IGFBP4, ILK, ITGA5, MAP1B, MITF, MMP2 (Gelatinase A), RGS2, SERPINE1 (PAI-1), SNAI1, SNAI2, SPARC, TCF4, TGFB1, TGFB2, TGFB3, TIMP1, TMEFF1, TSPAN13, VIM, VPS13A, WNT5A. Decreased H3K27me3: DSP, FGFBP1, GSC, IL1RN. Differentiation & Development: AKT1, BMP1, BMP2, BMP7, COL3A1, COL5A2, CTNNB1, DSP, ERBB3, F11R, FOXC2, FZD7, GSC, KRT14, MITF, MST1R, NODAL, NOTCH1, PTP4A1, SMAD2, SNAI1, SNAI2, SOX10, TGFB2, TGFB3, TMEFF1, TWIST1, VCAN, WNT11, WNT5A, WNT5B. Morphogenesis: CTNNB1, FOXC2, PPP3R1, RAC1, SMAD2, SNAI1, SOX10, TGFB1, TGFB2, TGFB3, TWIST1, WNT11, WNT5A. Cell Growth & Proliferation: AKT1, BMP1, BMP2, BMP7, CAV2, CTNNB1, EGFR, ERBB3, FGFBP1, FOXC2, HIF1A, IGFBP4, ILK, MST1R, NODAL, PDGFRB, TGFB1, TGFB2, TGFB3, TIMP1, VCAN. Migration & Motility: CALD1, CAV2, EGFR, FN1, ITGB1, MSN, MST1R, NODAL, PDGFRB, RAC1, STAT3, TGFB1, VIM. Cytoskeleton: CAV2, KRT7, MAP1B, PLEK2, RAC1, VIM. Extracellular Matrix & Cell Adhesion: BMP1, BMP2, BMP7, CDH1 (E-cadherin), CDH2 (N-cadherin), COL1A2, COL3A1, COL5A2, CTGF, CTNNB1, DSC2, EGFR, ERBB3, F11R, FN1, FOXC2, ILK, ITGA5, ITGAV, ITGB1, MMP2 (Gelatinase A), MMP3, MMP9 (Gelatinase B), PTK2, RAC1, SERPINE1 (PAI-1), SPP1 (Osteopontin), TGFB1, TGFB2, TIMP1, VCAN. Signaling Pathways: Estrogen Receptor: CAV2, ESR1 (ERa), KRT19, TGFB3. G-Protein Coupled Receptor: AKT1, FZD7, GNG11, RAC1, RGS2. Integrin-Mediated: COL3A1, CTGF, ILK, ITGA5, ITGAV, ITGB1, PTK2. Notch: FOXC2, NOTCH1. Receptor Tyrosine Kinase: EGFR, ERBB3, PDGFRB, RGS2, SPARC. TGFb / BMP: BMP1, BMP2, BMP7, COL3A1, SMAD2, SMAD4, TGFB1, TGFB2, TGFB3. WNT: CTNNB1, FZD7, GSK3B, WNT11, WNT5A, WNT5B. Transcription Factors: CTNNB1, ESR1 (ERa), FOXC2, GSC, MITF, NOTCH1, SIP1, SMAD2, SNAI2, SNAI3, SOX10, STAT3, TCF4, TWIST1, ZEB

18 miscript mirna PCR Arrays. (mirbase V19) Human Mouse Rat Dog Rhesus Macaque.miRNome (mirbase V19) 384 Well Human Mouse Rat Dog Rhesus Macaque.miFinder Human Mouse Rat Dog Rhesus Macaque.miFinder 384 Well Human Mouse.Apoptosis Human Mouse Rat.Brain Cancer Human Mouse Rat.Breast Cancer Human Mouse Rat.Cancer PathwayFinder Human Mouse Rat.Cardiovascular Disease Human Mouse Rat.Cell Differentiation & Development Human Mouse Rat.Immunopathology Human Mouse Rat.Inflammatory Response and Autoimmunity Human Mouse Rat.Neurological Development & Disease Human Mouse Rat.Ovarian Cancer Human Mouse Rat.Prostate Cancer Human Mouse Rat.Serum & Plasma Human Mouse Rat.Serum & Plasma 384 Well Human.T-Cell & B-Cell Activation Human Mouse Rat.miRNA QC Human Mouse Rat Dog Rhesus Macaque

19 mirna Targets PCR Arrays.After mirna expression analysis with miscript mirna PCR Arrays, perform the.following functional mirna study..once you have treated your cells with a mimic, inhibitor, or target protector, analyze target gene expression with the RT² Profiler mirna Targets PCR Arrays..A mirna target gene should meet the following criteria:.target gene expression inversely correlates with mirna expression.transfection with a miscript mirna Mimic downregulates target gene expression.transfection with a miscript mirna Inhibitor re-activates target gene expression.transfection with a miscript Target Protector re-activates target gene expression

20 ECM (Extracellular Matrix) and Cell Adhesion PCR Arrays Cell Adhesion Molecules: Transmembrane Molecules: CD44, CDH1, HAS1, ICAM1, ITGA1, ITGA2, ITGA3, ITGA4, ITGA5, ITGA6, ITGA7, ITGA8, ITGAL, ITGAM, ITGAV, ITGB1, ITGB2, ITGB3, ITGB4, ITGB5, MMP14, MMP15, MMP16, NCAM1, PECAM1, SELE, SELL, SELP, SGCE, SPG7, VCAM1. Cell-Cell Adhesion: CD44, CDH1, COL11A1, COL14A1, COL6A2, CTNND1, ICAM1, ITGA8, VCAM1. Cell-Matrix Adhesion: ADAMTS13, CD44, ITGA1, ITGA2, ITGA3, ITGA4, ITGA5, ITGA6, ITGA7, ITGA8, ITGAL, ITGAM, ITGAV, ITGB1, ITGB2, ITGB3, ITGB4, ITGB5, SGCE, SPP1, THBS3. Other Adhesion Molecules: CNTN1, COL12A1, COL15A1, COL16A1, COL5A1, COL6A1, COL7A1, COL8A1, VCAN, CTGF, CTNNA1, CTNNB1, CTNND2, FN1, KAL1, LAMA1, LAMA2, LAMA3, LAMB1, LAMB3, LAMC1, THBS1, THBS2, CLEC3B, TNC, VTN. Extracellular Matrix Proteins: Basement Membrane Constituents: COL4A2, COL7A1, LAMA1, LAMA2, LAMA3, LAMB1, LAMB3, LAMC1, SPARC. Collagens & ECM Structural Constituents: COL11A1, COL12A1, COL14A1, COL15A1, COL16A1, COL1A1, COL4A2, COL5A1, COL6A1, COL6A2, COL7A1, COL8A1, FN1, KAL1. ECM Proteases: ADAMTS1, ADAMTS13, ADAMTS8, MMP1, MMP10, MMP11, MMP12, MMP13, MMP14, MMP15, MMP16, MMP2, MMP3, MMP7, MMP8, MMP9, SPG7, TIMP1. ECM Protease Inhibitors: COL7A1, KAL1, THBS1, TIMP1, TIMP2, TIMP3. Other ECM Molecules: VCAN, CTGF, ECM1, HAS1, SPP1, TGFBI, THBS2, THBS3, CLEC3B, TNC, VTN Tsau C, Ito M, Gromova A, Hoffman MP, Meech R, Makarenkova HP. Barx2 and Fgf10 regulate ocular glands branching morphogenesis by controlling extracellular matrix remodeling. Development (2011) Aug;138(15):

21 Cell Junctions PCR Arrays There are 5 different types of cell junctions. They are tight junctions, adherens junctions, desmosomes, hemidesmosomes, and gap junctions. Focal Adhesions Tight Junctions Adherens Junctions Gap Junctions

22 Cancer Metastasis / Cell Motility PCR Arrays Cancer Metastasis PCR Array: 028Z.html Cell Motility PCR Array: Zhu W, Cai MY, Tong ZT, Dong SS, Mai SJ, Liao YJ, Bian XW, Lin MC, Kung HF, Zeng YX, Guan XY, Xie D. Overexpression of EIF5A2 promotes colorectal carcinoma cell aggressiveness by upregulating MTA1 through C- myc to induce epithelial-mesenchymaltransition. Gut (2012) Apr;61(4):

23 Fibrosis PCR Arrays Pro-Fibrotic: ACTA2 (a-sma), AGT, CCL11 (Eotaxin), CCL2 (MCP-1), CCL3 (MIP-1a), CTGF, GREM1, IL13, IL13RA2, IL4, IL5, SNAI1 (Snail). Anti-Fibrotic: BMP7, HGF, IFNG, IL10, IL13RA2. Extracellular Matrix & Cell Adhesion: ECM Components: COL1A2, COL3A1. Remodeling Enzymes: LOX, MMP1 (Collagenase 1), MMP13, MMP14, MMP2 (Gelatinase A), MMP3, MMP8, MMP9 (Gelatinase B), PLAT (tpa), PLAU (upa), PLG, SERPINA1 (a1-antitrypsin), SERPINE1 (PAI-1), SERPINH1, TIMP1, TIMP2, TIMP3, TIMP4. Cellular Adhesion: ITGA1, ITGA2, ITGA3, ITGAV, ITGB1, ITGB3, ITGB5, ITGB6, ITGB8. Inflammatory Cytokines & Chemokines: CCL11 (Eotaxin), CCL2 (MCP-1), CCL3 (MIP-1a), CCR2, CXCR4, IFNG, IL10, IL13, IL13RA2, IL1A, IL1B, IL4, IL5, ILK, TNF. Growth Factors: AGT, CTGF, EDN1, EGF, HGF, PDGFA, PDGFB, VEGFA. Signal Transduction: TGFß Superfamily: BMP7, CAV1, DCN, ENG (EVI-1), GREM1, INHBE, LTBP1, SMAD2, SMAD3, SMAD4, SMAD6, SMAD7, TGFB1, TGFB2, TGFB3, TGFBR1 (ALK5), TGFBR2, TGIF1, THBS1, THBS2 Transcription Factors: CEBPB, JUN, MYC, NFKB1, SP1, STAT1, STAT6 Epithelial-to-Mesenchymal Transition: AKT1, BMP7, COL1A2, COL3A1, ILK, ITGAV, ITGB1, MMP2 (Gelatinase A), MMP3, MMP9, SERPINE1 (PAI-1), SMAD2, SNAI1 (Snail), TGFB1, TGFB2, TGFB3, TIMP1. Others: BCL2, FASLG (TNFSF6)

24 Oxidative Stress PCR Arrays Antioxidants: Glutathione Peroxidases (GPx): GPX1, GPX2, GPX3, GPX4, GPX5, GPX6, GPX7, GSTP1, GSTZ1. Peroxiredoxins (TPx): PRDX1, PRDX2, PRDX3, PRDX4, PRDX5, PRDX6 (AOP2). Other Peroxidases: CAT, CYBB, CYGB, DUOX1, DUOX2, EPX, LPO, MGST3, MPO, PTGS1, PTGS2 (COX2), PXDN, TPO, TTN. Other Antioxidants: ALB, APOE, GSR, MT3, SELS, SOD1, SOD3, SRXN1, TXNRD1, TXNRD2. Genes Involved in Reactive Oxygen Species (ROS) Metabolism: Superoxide Dismutases (SOD): SOD1, SOD2, SOD3. Other Genes Involved in Superoxide Metabolism: ALOX12, CCS, DUOX1, DUOX2, GTF2I, MT3, NCF1, NCF2, NOS2 (inos), NOX4, NOX5, PREX1, UCP2. Other Genes Involved in ROS Metabolism: AOX1, BNIP3, EPHX2, MPV17, SFTPD. Oxidative Stress Responsive Genes: APOE, ATOX1, CAT, CCL5 (RANTES), CYGB, DHCR24, DUOX1, DUOX2, DUSP1 (PTPN16), EPX, FOXM1, FTH1, GCLC, GCLM, GPX1, GPX2, GPX3, GPX4, GPX5, GPX6, GPX7, GSR, GSS, HMOX1, HSPA1A, KRT1, LPO, MBL2, MPO, MSRA, NQO1, NUDT1, OXR1, OXSR1, PDLIM1, PNKP, PRDX2, PRDX5, PRDX6 (AOP2), PRNP, RNF7, SCARA3, SELS, SEPP1, SIRT2, SOD1, SOD2, SQSTM1, SRXN1, STK25, TPO, TTN, TXN, TXNRD1, TXNRD2. Oxygen Transporters: CYGB, MB Joyce NC, Harris DL, Zhu CC. Age-related gene response of human corneal endothelium to oxidative stress and DNA damage. Invest Ophthalmol Vis Sci (2011) Mar 1;52(3):

25 Hypoxia PCR Arrays. & Co-Transcription Factors: ARNT, COPS5, HIF1A, HIF1AN, HIF3A, HNF4A, NCOA1, PER1..Other HIF1 Interactors: APEX1, EGLN1, EGLN2, NFKB1, P4HA1, P4HB, TP53..Responsive Genes: Angiogenesis: ADORA2B, ANGPTL4, ANXA2, BTG1, EGR1, EDN1, EPO, F3, GPI, HMOX1, JMJD6, LOX, MMP9, PGF, PLAU (upa), SERPINE1 (PAI-1), VEGFA. Coagulation: ALDOA, ANXA2, F10, F3, F3, PLAU (upa), SERPINE1 (PAI-1), SLC16A3. DNA Damage Signaling & Repair: ATR, MIF, NDRG1, RUVBL2. Metabolism: ALDOA, DDIT4 (REDD1), ENO1, ERO1L, GBE1, GPI, GYS1, HK2, LDHA, PDK1, PFKFB3, PFKFB4, PFKL, PFKP, PGAM1, PGK1, PKM2, SLC2A1, SLC2A3, TPI1. Regulation of Apoptosis: ADM, BNIP3, BNIP3L, BTG1, DDIT4 (REDD1), IER3, MIF, NOS3 (enos), PIM1. Regulation of Cell Proliferation: ADM, BTG1, BLM, CCNG2, EGR1, IGFBP3, MET, MIF, MXI1, NAMPT, NOS3 (enos), ODC1, PGF, PIM1, TXNIP. Transcription Factors: BHLHE40, FOS, RBPJ, USF2. Transporters, Channels & Receptors: SLC2A1, SLC2A3, SLC16A3, TFRC, VDAC1. Other Responsive Genes: ANKRD37, CA9, CTSA, DNAJC5, EIF4EBP1, LGALS3, MAP3K Mueller BR, Bale TL. Sex-specific programming of offspring emotionality after stress early in pregnancy. J Neurosci (2008) Sep 3;28(36):

26 NFκB Signaling or Targets PCR Arrays NFκB Signaling Pathway PCR Array: NFκB Signaling Targets PCR Array: Fiume G, Vecchio E, De Laurentiis A, Trimboli F, Palmieri C, Pisano A, Falcone C, Pontoriero M, Rossi A, Scialdone A, Fasanella Masci F, Scala G, Quinto I. Human immunodeficiency virus-1 Tat activates NF-kappaB via physical interaction with IkappaB-alpha and p65. Nucleic Acids Res Dec

27 TGFβ Signaling or Targets PCR Arrays TGFβ / BMP Signaling Pathway PCR Array: TGFβ Signaling Targets PCR Array: Garamszegi N, Garamszegi SP, Shehadeh LA, Scully SP. Extracellular matrix-induced gene expression in human breast cancer cells. Mol Cancer Res. (2009) Mar;7(3):

28 Wnt Signaling or Targets PCR Arrays Wnt Signaling Pathway PCR Array: Wnt Signaling Targets PCR Array: Burkhalter RJ, Symowicz J, Hudson LG, Gottardi CJ, Stack MS. Integrin regulation of beta-catenin signaling in ovarian carcinoma. J Biol Chem. (2011) Jul 1;286(26): Hussain M, Rao M, Humphries AE, Hong JA, Liu F, Yang M, Caragacianu D, Schrump DS. Tobacco smoke induces polycombmediated repression of Dickkopf-1 in lung cancer cells. Cancer Res. (2009) Apr 15;69(8):

29 Contact Information.Jesse Liang. Support: Webinar Calendar:

Supplementary Table 1: List of the 242 hypoxia/reoxygenation marker genes collected from literature and databases

Supplementary Tables: Supplementary Table 1: List of the 242 hypoxia/reoxygenation marker genes collected from literature and databases Supplementary Table 2: Contingency tables used in the fisher exact