Supplemental Fig. 1 12 1mg/ml A1AT p<0.001 10 8 p=0.004 6 p=0.029 (fold change) 4 mRNA ofmRNA angptl4 at h 1 2 0 Control Prolastin Aralast A1AT Sigma Supplemental Figure 1. Adherent PBMCs were incubated with 1 mg/ml A1AT from different sources (Prolastin, Grifols; Aralast, Baxter; or Sigma Aldrich) for 1h and mRNA levels of angplt4 were determined by RT-PCR. Bars represent the mean ± SD of four experiments. Supplemental Fig. 2 12 8 h p=0.005 10 8 6 mRNA angplt4 mRNA change Fold 4 2 0 Control A1AT 0.5 mg/ml Supplemental Figure 2. HMVEC-L were incubated with 0.5 mg/ml A1AT (Calbiochem) for 8 h and mRNA levels of angptl4 were determined by RT-PCR. Bars represent the mean ± SEM of three experiments. Supplemental Table I. Influence of A1AT (1 mg/ml, 8 h) on expression of angiogenesis genesa in HMVEC-L Unigene Symbol Description Gname fold changeb Hs.525622 AKT1 V-akt murine thymoma viral AKT/MGC99656/PKB/PKB- 0,94 0,14 oncogene homolog 1 ALPHA/PRKBA/RAC/RAC- ALPHA Hs.369675 ANGPT1 Angiopoietin 1 AGP1/AGPT/ANG1 3,61 5,11 Hs.583870 ANGPT2 Angiopoietin 2 AGPT2/ANG2 0,98 0,01 Hs.209153 ANGPTL3 Angiopoietin-like 3 ANGPT5/FHBL2 1,10 0,59 Hs.9613 ANGPTL4 Angiopoietin-like 4 ANGPTL2/ARP4/FIAF/HFAR 5,79 1,66 P/NL2/PGAR/pp1158 Hs.1239 ANPEP Alanyl (membrane) APN/CD13/GP150/LAP1/P150 1,03 0,01 aminopeptidase /PEPN Hs.194654 BAI1 Brain-specific angiogenesis FLJ41988/GDAIF 0,89 0,26 inhibitor 1 Hs.54460 CCL11 Chemokine (C-C motif) ligand 11 MGC22554/SCYA11 1,04 0,15 Hs.303649 CCL2 Chemokine (C-C motif) ligand 2 GDCF- 1,09 0,14 2/HC11/HSMCR30/MCAF/M CP- 1/MCP1/MGC9434/SCYA2/S MC-CF Hs.76206 CDH5 Cadherin 5, type 2 (vascular 7B4/CD144/FLJ17376 0,96 0,14 endothelium) Hs.517356 COL18A1 Collagen, type XVIII, alpha 1 FLJ27325/FLJ34914/KNO/KN 0,99 0,09 O1/KS/MGC74745 Hs.570065 COL4A3 Collagen, type IV, alpha 3 - 0,62 0,51 (Goodpasture antigen) Hs.789 CXCL1 Chemokine (C-X-C motif) ligand 1 FSP/GRO1/GROa/MGSA/MG 0,93 0,09 (melanoma growth stimulating SA-a/NAP-3/SCYB1 activity, alpha) Hs.632586 CXCL10 Chemokine (C-X-C motif) ligand C7/IFI10/INP10/IP- 1,49 0,56 10 10/SCYB10/crg-2/gIP-10/mob- 1 Hs.89690 CXCL3 Chemokine (C-X-C motif) ligand 3 CINC-2b/GRO3/GROg/MIP- 0,66 0,23 2b/MIP2B/SCYB3 Hs.89714 CXCL5 Chemokine (C-X-C motif) ligand 5 ENA-78/SCYB5 1,54 0,36 Hs.164021 CXCL6 Chemokine (C-X-C motif) ligand 6 CKA-3/GCP-2/GCP2/SCYB6 0,90 0,13 (granulocyte chemotactic protein 2) Hs.77367 CXCL9 Chemokine (C-X-C motif) ligand 9 CMK/Humig/MIG/SCYB9/crg 0,45 0,33 -10 Hs.592212 TYMP Thymidine phosphorylase ECGF/ECGF1/MEDPS1/MNG 1,26 0,06 IE/MTDPS1/PDECGF/TP/hPD -ECGF Hs.154210 S1PR1 Sphingosine-1-phosphate receptor CHEDG1/D1S3362/ECGF1/E 1,02 0,25 1 DG-1/EDG1/FLJ58121/S1P1 Hs.516664 EFNA1 Ephrin-A1 B61/ECKLG/EFL1/EPLG1/LE 0,80 0,11 RK1/TNFAIP4 Hs.516656 EFNA3 Ephrin-A3 EFL2/EPLG3/Ehk1-L/LERK3 0,87 0,23 Hs.149239 EFNB2 Ephrin-B2 EPLG5/HTKL/Htk- 0,79 0,17 L/LERK5/MGC126226/MGC1 26227/MGC126228 Hs.419815 EGF Epidermal growth factor HOMG4/URG 1,01 0,36 Hs.76753 ENG Endoglin CD105/END/FLJ41744/HHT1/ 1,07 0,01 ORW/ORW1 Hs.437008 EPHB4 EPH receptor B4 HTK/MYK1/TYRO11 0,89 0,13 Hs.115263 EREG Epiregulin ER 1,50 0,10 Hs.483635 FGF1 Fibroblast growth factor 1 (acidic) AFGF/ECGF/ECGF- 0,89 0,55 beta/ECGFA/ECGFB/FGF- alpha/FGFA/GLIO703/HBGF1 Hs.284244 FGF2 Fibroblast growth factor 2 (basic) BFGF/FGFB/HBGF-2 1,01 0,17 Hs.1420 FGFR3 Fibroblast growth factor receptor 3 ACH/CD333/CEK2/HSFGFR3 1,36 0,29 EX/JTK4 Hs.11392 FIGF C-fos induced growth factor VEGF-D/VEGFD 1,37 0,29 (vascular endothelial growth factor D) Hs.654360 FLT1 Fms-related tyrosine kinase 1 FLT/VEGFR1 1,10 0,14 (vascular endothelial growth factor/vascular permeability factor receptor) Hs.388245 HAND2 Heart and neural crest derivatives DHAND2/FLJ16260/Hed/MG 3,13 3,75 expressed 2 C125303/MGC125304/Thing2/ bHLHa26/dHand Hs.396530 HGF Hepatocyte growth factor DFNB39/F- 1,12 0,38 (hepapoietin A; scatter factor) TCF/HGFB/HPTA/SF Hs.597216 HIF1A Hypoxia inducible factor 1, alpha HIF-1alpha/HIF1/HIF1- 1,12 0,07 subunit (basic helix-loop-helix ALPHA/MOP1/PASD8/bHLH transcription factor) e78 Hs.44227 HPSE Heparanase HPA/HPA1/HPR1/HPSE1/HS 1,04 0,12 E1 Hs.504609 ID1 Inhibitor of DNA binding 1, ID/bHLHb24 0,71 0,18 dominant negative helix-loop-helix protein Hs.76884 ID3 Inhibitor of DNA binding 3, HEIR-1/bHLHb25 0,76 0,16 dominant negative helix-loop-helix protein Hs.37026 IFNA1 Interferon, alpha 1 IFL/IFN/IFN-ALPHA/IFN- 1,42 1,14 alphaD/IFNA13/IFNA@/MGC 138207/MGC138505/MGC138 507 Hs.93177 IFNB1 Interferon, beta 1, fibroblast IFB/IFF/IFNB/MGC96956 2,70 1,76 Hs.856 IFNG Interferon, gamma IFG/IFI 1,05 0,14 Hs.160562 IGF1 Insulin-like growth factor 1 IGF-I/IGF1A/IGFI 1,22 0,15 (somatomedin C) Hs.126256 IL1B Interleukin 1, beta IL-1/IL1-BETA/IL1F2 1,52 0,41 Hs.654458 IL6 Interleukin 6 (interferon, beta 2) BSF2/HGF/HSF/IFNB2/IL-6 0,89 0,20 Hs.624 IL8 Interleukin 8 CXCL8/GCP- 1,10 0,21 1/GCP1/LECT/LUCT/LYNAP /MDNCF/MONAP/NAF/NAP- 1/NAP1 Hs.436873 ITGAV Integrin, alpha V (vitronectin CD51/DKFZp686A08142/MS 1,14 0,14 receptor, alpha polypeptide, K8/VNRA antigen CD51) Hs.218040 ITGB3 Integrin, beta 3 (platelet CD61/GP3A/GPIIIa 1,07 0,09 glycoprotein IIIa, antigen CD61) Hs.728907 JAG1 Jagged 1 AGS/AHD/AWS/CD339/HJ1/J 1,36 0,30 AGL1/MGC104644 Hs.479756 KDR Kinase insert domain receptor (a CD309/FLK1/VEGFR/VEGFR 0,96 0,12 type III receptor tyrosine kinase) 2 Hs.473256 LAMA5 Laminin, alpha 5 KIAA1907 1,11 0,19 Hs.421391 LECT1 Leukocyte cell derived chemotaxin BRICD3/CHM- 1,92 2,26 1 I/CHM1/MYETS1 Hs.194236 LEP Leptin FLJ94114/OB/OBS 1,05 0,14 Hs.82045 MDK Midkine (neurite growth- FLJ27379/MK/NEGF2 0,92 0,19 promoting factor 2) Hs.513617 MMP2 Matrix metallopeptidase 2 CLG4/CLG4A/MMP- 0,95 0,08 (gelatinase A, 72kDa gelatinase, II/MONA/TBE-1 72kDa type IV collagenase) Hs.297413 MMP9 Matrix metallopeptidase 9 CLG4B/GELB/MANDP2/MM 1,03 0,52 (gelatinase B, 92kDa gelatinase, P-9 92kDa type IV collagenase) Hs.436100 NOTCH4 Notch 4 FLJ16302/INT3/MGC74442/N 0,96 0,08 OTCH3 Hs.131704 NRP1 Neuropilin 1 BDCA4/CD304/DKFZp686A0 0,98 0,16 3134/DKFZp781F1414/NP1/N RP/VEGF165R Hs.471200 NRP2 Neuropilin 2 MGC126574/NP2/NPN2/PRO 1,08 0,18 2714/VEGF165R2 Hs.535898 PDGFA Platelet-derived growth factor PDGF-A/PDGF1 1,11 0,06 alpha polypeptide Hs.514412 PECAM1 Platelet/endothelial cell adhesion CD31/FLJ34100/FLJ58394/PE 1,10 0,29 molecule CAM-1 Hs.81564 PF4 Platelet factor 4 CXCL4/MGC138298/SCYB4 0,72 0,30 Hs.252820 PGF Placental growth factor D12S1900/PGFL/PLGF/PlGF- 1,06 0,21 2/SHGC-10760 Hs.77274 PLAU Plasminogen activator, urokinase ATF/UPA/URK/u-PA 1,12 0,13 Hs.143436 PLG Plasminogen DKFZp779M0222 0,76 0,35 Hs.125036 PLXDC1 Plexin domain containing 1 DKFZp686F0937/FLJ36270/F 0,49 0,13 LJ45632/TEM3/TEM7 Hs.528665 PROK2 Prokineticin 2 BV8/KAL4/MIT1/PK2 1,03 0,24 Hs.201978 PTGS1 Prostaglandin-endoperoxide COX1/COX3/PCOX1/PGG/H 0,73 0,12 synthase 1 (prostaglandin G/H S/PGHS- synthase and cyclooxygenase) 1/PGHS1/PHS1/PTGHS Hs.532768 SERPINF1 Serpin peptidase inhibitor, clade F EPC-1/PEDF 1,08 0,12 (alpha-2 antiplasmin, pigment epithelium derived factor), member 1 Hs.68061 SPHK1 Sphingosine kinase 1 SPHK 0,98 0,16 Hs.301989 STAB1 Stabilin 1 CLEVER-1/FEEL-1/FELE- 1,03 0,08 1/FEX1/KIAA0246/STAB-1 Hs.89640 TEK TEK tyrosine kinase, endothelial CD202B/TIE- 0,91 0,04 2/TIE2/VMCM/VMCM1 Hs.170009 TGFA Transforming growth factor, alpha TFGA 0,82 0,29 Hs.645227 TGFB1 Transforming growth factor, beta 1 CED/DPD1/LAP/TGFB/TGFb 0,96 0,02 eta Hs.133379 TGFB2 Transforming growth factor, beta 2 MGC116892/TGF-beta2 0,90 0,14 Hs.494622 TGFBR1 Transforming growth factor, beta AAT5/ACVRLK4/ALK- 1,28 0,03 receptor 1 5/ALK5/LDS1A/LDS2A/SKR 4/TGFR-1 Hs.164226 THBS1 Thrombospondin 1 THBS/THBS-1/TSP/TSP- 1,09 0,15 1/TSP1 Hs.371147 THBS2 Thrombospondin 2 TSP2 0,98 0,06 Hs.522632 TIMP1 TIMP metallopeptidase inhibitor 1 CLGI/EPA/EPO/FLJ90373/HC 1,01 0,14 I/TIMP Hs.633514 TIMP2 TIMP metallopeptidase inhibitor 2 CSC-21K 0,88 0,05 Hs.644633 TIMP3 TIMP metallopeptidase inhibitor 3 HSMRK222/K222/K222TA2/S 2,79 2,95 FD Hs.241570 TNF Tumor necrosis factor DIF/TNF- 1,06 0,10 alpha/TNFA/TNFSF2 Hs.525607 TNFAIP2 Tumor necrosis factor, alpha- B94/EXOC3L3 1,19 0,12 induced protein 2 Hs.73793 VEGFA Vascular endothelial growth factor MGC70609/MVCD1/VEGF/V 1,02 0,34 A PF Hs.435215 VEGFC Vascular endothelial growth factor Flt4-L/VRP 1,10 0,17 C Hs.534255 B2M Beta-2-microglobulin - 0,99 0,08 Hs.412707 HPRT1 Hypoxanthine HGPRT/HPRT 1,04 0,14 phosphoribosyltransferase 1 Hs.728776 RPL13A Ribosomal protein L13a L13A/TSTA1 0,96 0,03 Hs.592355 GAPDH Glyceraldehyde-3-phosphate G3PD/GAPD/MGC88685 0,93 0,05 dehydrogenase Hs.520640 ACTB Actin, beta PS1TP5BP1 1,10 0,12 aGene expression was analyzed using angiogenesis PCR arrays.