Supplementary Table 1. Composition of the diets
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- Ἰωνᾶς Κοσμόπουλος
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1 Supplementary Table 1. Composition of the diets Standard diet High Fat diet Energy content (kcal/kg) Protein (g/kg) Carbohydrate (g/kg) Total fat (g/kg) Saturated fatty acids Capric acid (g/kg) 4.7 Lauric acid (g/kg) 33.4 Myristic acid (g/kg) Palmitic acid (g/kg) Stearic acid (g/kg) Arachidic acid (g/kg) Unsaturated fatty acids Palmitoleic acid (g/kg) Oleic acid (g/kg) Linoleic acid (g/kg) Linolenic acid (g/kg) Eicosaenoic acid (g/kg) Arachidonic acid (g/kg) 0 0.4
2 Supplementary Table 2. List of Antibodies Primary Antibody Species Dilution Manufacturer Corresponding Secondary Antibody & Dilution Akt Rabbit 1:1000 Cell Signaling HRP anti rabbit, 1:2500 Activin A receptor type 1 B (Alk4) Rabbit 1:1000 Abcam HRP anti rabbit, 1:2500 Calreticulin Rabbit 1:1000 Stressgen Bioreagents HRP anti rabbit, 1:2500 GAPDH Rabbit 1:10000 Abcam HRP anti rabbit, 1:2500 Glut 4 (C 20) Goat 1:1000 Santa Cruz p Akt Ser473 Rabbit 1:1000 Cell Signaling p Phospholamban Mouse 1:200 Santa Cruz HRP anti goat, 1:2500 HRP anti rabbit, 1:2500 HRP anti mouse, 1:2500 p Smad Ser465/467 Rabbit 1:500 Cell Signaling HRP anti rabbit, 1:2500 Serca2a Mouse 1:1000 Abcam HRP anti mouse, 1:2500 SMAD2/3 1:1000 B&D Bioscience HRP anti mouse, 1:2500 α Tubulin Mouse 1:1000 Calbiochem HRP anti mouse, 1:2500
3 Supplementary Table 3. Immunoreactivity in conditioned medium collected from epicardial and subcutaneous adipose tissue explants from guinea pigs Swiss Prot Name on array Symbol (n=7) (n=7) Q15848 Acrp30 ADIPOQ 182 ± ± 166 P08476 Activin A INHBA 1718 ± ± 212 O00253 AgRP AGRP 1434 ± ± 141 Q13740 ALCAM ALCAM 8149 ± ± 621 P15514 Amphiregulin AREG 1998 ± ± 557 P03950 Angiogenin ANG 208 ± ± 64 O15123 Angiopoietin 2 ANGPT ± ± 342 P30530 Axl AXL 712 ± ± 65 P33681 B7 1 (CD80) CD ± ± 83 P23560 BDNF BDNF 820 ± ± 144 P09038 bfgf FGF ± ± 498 O43927 BLC CXCL ± ± 74 P12644 BMP 4 BMP4 719 ± ± 227 P22003 BMP 5 BMP ± ± 300 P22004 BMP 6 BMP ± ± 297 P18075 BMP 7 BMP7 358 ± ± 80 P01138 b NGF NGF 4372 ± ± 723 P35070 BTC BTC 1037 ± ± 127 Q16619 Cardiotrophin 1 CTF1 941 ± ± 88 Q9NRJ3 CCL 28 CCL ± ± 72 P08571 CD14 CD ± ± 58 P55773 CK b 8 1 CCL ± ± 246 P26441 CNTF CNTF 459 ± ± 61 Q9Y4X3 CTACK CCL ± ± 314 Q9H2A7 CXCL 16 CXCL ± ± 222 O75509 DR6 (TNFRSF21) TNFRSF ± ± 138 Q06418 Dtk TYRO3 637 ± ± 179 P01133 EGF EGF 730 ± ± 235 P00533 EGF R EGFR 347 ± ± 176 P42830 ENA 78 CXCL5 966 ± ± 313 P17813 Endoglin ENG 515 ± ± 77 P51671 Eotaxin CCL ± ± 138 O00175 Eotaxin 2 CCL ± ± 295 Q9Y258 Eotaxin 3 CCL ± ± 146 P21860 ErbB3 ERBB3 834 ± ± 88 P16581 E Selectin SELE 440 ± ± 85 P48023 Fas Ligand FASLG 843 ± ± 143 Q549F0 Fas/TNFRSF6 FAF ± ± 856 P08620 FGF 4 FGF ± ± 1427 P10767 FGF 6 FGF6 283 ± ± 95 P21781 FGF 7 FGF7 273 ± ± 65 P31371 FGF 9 FGF9 846 ± ± 151 P49771 Fit 3 Ligand FLT3LG 601 ± ± 242 P78423 Fractalkine CX3CL1 302 ± ± 63
4 P80162 GCP 2 CXCL6 339 ± ± 79 P09919 GCSF CSF3 513 ± ± 246 P39905 GDNF GDNF 3799 ± ± 1646 Q9Y5U5 GITR TNFRSF ± ± 1605 Q9UNG2 GITR Ligand TNFSF ± ± 466 P04141 GM CSF CSF2 196 ± ± 44 P09341 GRO CXCL1 699 ± ± 94 P09341 GROα CXCL1 217 ± ± 107 O15467 HCC 4 CCL ± ± 423 P14210 HGF HGF 254 ± ± 63 P22362 I 309 CCL1 292 ± ± 110 P05362 ICAM 1 ICAM1 521 ± ± 108 P13598 ICAM 2 ICAM ± ± 3916 P32942 ICAM 3 ICAM3 703 ± ± 136 P01343 IGF 1 IGF1 443 ± ± 49 P08833 IGFBP 1 IGFBP1 531 ± ± 136 P18065 IGFBP 2 IGFBP2 348 ± ± 85 P17936 IGFBP 3 IGFBP3 794 ± ± 579 P22692 IGFBP 4 IGFBP4 582 ± ± 145 P24592 IGFBP 6 IGFBP ± ± 246 P08069 IGF I SR IGF1R 290 ± ± 93 P01344 IGF II IGF ± ± 156 P27930 IL 1 R II IL1R ± ± 1867 Q01638 IL 1 RA/ST2 IL1RL1 683 ± ± 270 P14778 IL 1 RI IL1R1 362 ± ± 64 P22301 IL 10 IL ± ± 138 Q08334 IL 10 Rβ IL10RB 352 ± ± 115 P20809 IL 11 IL ± ± 243 P29459 IL 12 p40 IL12A 893 ± ± 211 P29460 IL 12 p70 IL12B 695 ± ± 431 P35225 IL 13 IL ± ± 78 Q14627 IL 13 Rα2 IL13RA2 847 ± ± 132 P40933 IL 15 IL ± ± 57 Q14005 IL 16 IL ± ± 97 Q16552 IL 17 IL17A 422 ± ± 354 Q13478 IL 18 BPα IL18R1 224 ± ± 37 O95256 IL 18 Rβ IL18RAP 802 ± ± 90 P01583 IL 1a IL1A 670 ± ± 215 P01584 IL 1b IL1B 192 ± ± 60 P18510 IL 1ra IL1RN 191 ± ± 75 P60568 IL 2 IL2 114 ± ± 20 P31785 IL 2 R IL2RG 606 ± ± 123 P01589 IL 2 Rα IL2RA 493 ± ± 38 P14784 IL 2 Rβ IL2RB 715 ± ± 482 Q9HBE5 IL 21R IL21R 614 ± ± 54 P08700 IL 3 IL3 178 ± ± 58 P05112 IL 4 IL4 264 ± ± 186 P05113 IL 5 IL5 201 ± ± 38
5 Q01344 IL 5 Rα IL5RA 514 ± ± 52 P05231 IL 6 IL6 519 ± ± 96 P08887 IL 6R IL6R 1185 ± ± 214 P13232 IL 7 IL7 150 ± ± 81 P10145 IL 8 IL8 420 ± ± 209 P15248 IL 9 IL9 831 ± ± 188 P01579 INF IFNG 168 ± ± 47 P02778 IP 10 CXCL ± ± 123 O14625 I TAC CXCL ± ± 133 P17676 LAP CEBPB 693 ± ± 98 P41159 Leptin LEP 491 ± ± 89 P48357 Leptin R LEPR 838 ± ± 167 P15018 LIF LIF 646 ± ± 159 O43557 LIGHT TNFSF ± ± 183 P14151 L Selectin SELL 511 ± ± 139 P47992 Lymphotactin XCL ± ± 476 P13500 MCP 1 CCL2 304 ± ± 64 P80075 MCP 2 CCL8 454 ± ± 73 P80098 MCP 3 CCL7 218 ± ± 74 Q99616 MCP 4 CCL ± ± 44 P09603 M CSF CSF1 498 ± ± 191 P07333 M CSF R CSF1R 376 ± ± 52 O00626 MDC CCL ± ± 224 P14174 MIF MIF 2203 ± ± 1565 Q07325 MIG CXCL9 379 ± ± 112 P10147 MIP 1a CCL ± ± 115 P13236 MIP 1b CCL ± ± 539 Q16663 MIP 1d CCL ± ± 199 P78556 MIP 3a CCL ± ± 90 Q99731 MIP 3b CCL ± ± 62 P03956 MMP 1 MMP ± ± 276 P45452 MMP 13 MMP ± ± 76 P08254 MMP 3 MMP3 478 ± ± 73 P14780 MMP 9 MMP9 406 ± ± 63 P55773 MPIF 1 CCL ± ± 35 P26927 MSPa MST1 559 ± ± 131 P02775 NAP 2 PPBP 3162 ± ± 1257 P08138 NGF R NGFR 272 ± ± 20 P20783 NT 3 NTF3 594 ± ± 182 P34130 NT 4 NTF4 617 ± ± 116 P13725 Oncostatin OSM 575 ± ± 91 O00300 Osteoprotegerin TNFRSF11B 552 ± ± 119 P55774 PARC CCL ± ± 322 P04085 PDGF AA PDGFA 897 ± ± 149 P16234 PDGF Rα PDGFRA 395 ± ± 44 P09619 PDGF Rβ PDGFRB 386 ± ± 182 na PDGF AB 661 ± ± 77 P01127 PDGF BB PDGFB 479 ± ± 118
6 P16284 PECAM 1 PECAM1 278 ± ± 48 Q07326 PIGF PIGF 982 ± ± 286 P01236 Prolactin PRL 554 ± ± 85 P13501 RANTES CCL ± ± 885 P21583 SCF KITLG 563 ± ± 93 P10721 SCF R KIT 679 ± ± 87 P48061 SDF 1 CXCL ± ± 207 P48061 SDF 1β CXCL ± ± 45 P40189 sgp 130 IL6ST 1782 ± ± 370 O15389 Siglec 5 SIGLEC5 406 ± ± 54 P19438 stnfr I TNFRSF1A 2271 ± ± 1287 P20333 stnfr II TNFRSF1B 1194 ± ± 288 Q92583 TARC CCL ± ± 36 O15444 TECK CCL ± ± 149 P01135 TGFα TGFA 728 ± ± 222 P01137 TGFβ1 TGFB1 916 ± ± 397 P61812 TGFβ2 TGFB ± ± 364 P10600 TGFβ3 TGFB3 398 ± ± 31 P40225 Thrombopoeitin THPO 729 ± ± 85 P35590 Tie 1 TIE1 516 ± ± 119 Q02763 Tie 2 TEK 734 ± ± 90 P01033 TIMP 1 TIMP1 882 ± ± 553 P16035 TIMP 2 TIMP ± ± Q99727 TIMP 4 TIMP ± ± 3623 P01375 TNFα TNF 248 ± ± 66 P01374 TNFβ LTA 814 ± ± 179 O14798 TRAIL R3 TNFRSF10C 361 ± ± 87 Q9UBN6 TRAIL R4 TNFRSF10D 628 ± ± 86 Q03405 upar PLAUR 450 ± ± 102 P33151 VE Cadherin CDH5 437 ± ± 111 P15692 VEGF VEGFA 727 ± ± 723 P35968 VEGF R2 KDR 536 ± ± 54 P35916 VEGF R3 FLT ± ± 1781 O43915 VEGF D FIGF 2536 ± ± 1816, P<5 epicardial adipose tissue () versus subcutaneous adipose tissue ()
7 A B shortening rate (µm/s) s ning (µm) sarcomere shorte C relengthening rate (µm m/s) Supplementary Figure 1. Effect of conditioned medium from epicardial and subcutaneous adipose tissue Supplementary Figure 1. Effect of conditioned medium from epicardial and subcutaneous adipose tissue on sarcomere shortening in cardiomyocytes. Primary adult rat cardiomyocytes were incubated with control AM or CM (diluted 1:2) from and from SD and HFD fed guinea pigs for 30 min before analysis of contractile function. A C. Effect of exposure of cardiomyocytes to CM on departure velocity of contraction (A), peak sarcomere shortening (B), and return velocity of contraction (C). Open bars, AM; gray bars, CM from SD fed animals; black bars, CM from HFD fedanimals.dataareexpressedasmean±s.e.m. of at least 8 independent experiments. Differences between the experimental groups were calculated by one way ANOVA and unpaired students t tests., P<5 versus AM;, P<5, HFD versus SD;, P<5 versus.
8 A B Ca 2+ increa ase (ratio/s) peak [Ca 2+ ] (ratio) C 00 Ca 2+ decrease (ratio/s) Supplementary Figure 2. Effect of conditioned medium from epicardial and subcutaneous adipose tissue on cytosolic [Ca 2+ ]incardiomyocytes.primary adult rat cardiomyocytes were incubated with control AM or CM (diluted 1:2) from and from SD and HFD fed guinea pigs for 30 min before analysis of cytosolic [Ca 2+ ]. A C. Effect of exposure of cardiomyocytes to CM on [Ca 2+ ]increase(a), peak [Ca 2+] (B), and [Ca 2+ ]decrease(c). Open bars, AM; gray bars, CM from SD fed animals; black bars, CM from HFD fed animals. Data are expressed as mean ± S.E.M. of at least 8 independent experiments. Differences between the experimental groups were calculated by one way ANOVA and unpaired students t tests., P<5 versus AM;, P<5, HFD versus SD;, P<5 versus.
9 A shortening rate (µm m/s) D Ca 2+ inc rease (ratio/s) B sarcomere shortening (µm) E peak [C Ca 2+ ] (ratio) C 2.0 F relenghtenin ng rate (µm/s) Ca 2+ decrease (ratio/ /s) Supplementary Figure 3. Contractile function in cardiomyocytes 2 hours after removal of conditioned media. Primary adult rat cardiomyocytes were incubated with control AM or CM from and from SD and HFD fed guinea pigs for 30 min. Then, the CM were replaced by control AM. Following a 2 h incubation, contractile function and cytosolic [Ca 2+ ] was analyzed. A C. Reversibility effect of exposure of cardiomyocytes to CM on departure velocity of contraction (A), peak sarcomere shortening (B), and return velocity of contraction (C). D F. Reversibility effect of exposure of cardiomyocytes to CM on [Ca 2+ ] increase (D), peak [Ca 2+ ](E), and [Ca 2+ ]decrease(f). Black bars, AM; gray bars, cells exposed to CM diluted 1:4; open bars, cells exposed to CM diluted 1:4. Data are mean ± S.E.M. from at least 8 independent experiments. Groups were compared by one way ANOVA and unpaired students t test., P<5 versus AM;, P<5, HFD versus SD.
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和光純薬工業の サイトカイン グロースファクター 各製詳細は 検索サイト Siyaku.com ( http://www.siyaku.com/ ) をご覧下さい 大入り包装の価格 納期は当社代理店までお問合せ下さい Human, recombinant 014-23961 018-23964 014-19671 016-23781 019-20731 013-20251 010-20261 017-20271
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