Published October 15, 2012 Supplemental material Hameed et al. http://www.jem.org/cgi/content/full/jem.20112716/DC1 Clinical details for human cells (Fig. 1, A and G) Sample Diagnosis Gender Age mPAP CO 1 2 3 4 5 6 IPAH IPAH hPAH (R491M BMPR2) Emphysema Centrilobular Emphysema Squamous cell carcinoma Female Female Female Female Male Female yr 38 23 45 58 62 60 mm Hg 56 70 50 n/a n/a n/a liters/min 2.8 2.0 2.4 n/a n/a n/a IPAH, idiopathic PAH; hPAH, hereditary PAH; CO, cardiac output; n/a, not available. Table S2. Hemodynamic data from MCT rats procedures Phenotype parameter Control day 21 MCT day 21 MCT day 21 (prevention) +IgG Heart rate (BPM) Cardiac index (µl/min/g) RVSP (mm Hg) RVEDP (mm Hg) RV dP/dtmax (mm Hg/s) RV dP/dtmin (mm Hg/s) LVESP (mm Hg) LVEDP (mm Hg) LV dP/dtmax (mm Hg/s) LV dP/dtmin (mm Hg/s) Mean aortic pressure (mm Hg) ePVRi (mm Hg/ml/min/g) PA AT (ms) RV/LV&Septum 319 ± 16 136 ± 26 25 ± 4.4 4±1 1,225 ± 142 1,339 ± 274 102 ± 5 4±1 6,876 ± 518 8,299 ± 670 96 ± 10 73 ± 22 34 ± 2 0.28 ± 0.001 331 ± 15 128 ± 42 50 ± 8 9±3 2,951 ± 422 2,296 ± 224 97 ± 6 6±1 5,428 ± 810 4,704 ± 283 85.6 ± 7 137 ± 52 22 ± 5 0.48 ± 0.03 298 ± 18 145 ± 24 43 ± 3 9±1 2,188 ± 192 1,995 ± 404 124 ± 4 5±1 7,378 ± 684 9,315 ± 841 102 ± 5 193 ± 31 22 ± 3 0.39 ± 0.05 +TRAIL Ab 299 ± 10 178 ± 12 23 ± 2** 5±2 1,193 ± 183 919 ± 157 125 ± 7 6±1 7,248 ± 470 7473 ± 622 96 ± 7 57 ± 11** 30 ± 3* 0.30 ± 0.01 MCT day 35 (survival) +IgG +TRAIL Ab 279 ± 21 281 ± 17 116 ± 15 147 ± 17 83 ± 2 90 ± 14 7±1 6±1 3,765 ± 573 4,818 ± 529 2,452 ± 254 4,178 ± 1479 92 ± 11 96 ± 9 5±1 4±1 5,466 ± 899 4,887 ± 870 4,565 ± 1144 3,581 ± 426 72 ± 9 96 ± 5 422 ± 38 339 ± 67 12 ± 1 12 ± 1 0.47 ± 0.1 0.52± 0.05 Data represent mean ± SEM. *, P < 0.05; **, P < 0.01, anti-TRAIL antibody versus IgG with each genotype as determined by Mann-Whitney U test. RV/LV&Septum, right ventricle/left ventricle including septum. JEM S1 Downloaded from jem.rupress.org on October 22, 2012 The Jour nal of Exper imental Medicine Table S1. Published October 15, 2012 Table S3. Hemodynamic data from TRAIL/ mouse hypoxia procedures Phenotype parameter C57BL/6 Normoxia Hypoxia 352 ± 8 134 ± 8 23 ± 1 3±1 1,603 ± 178 1,924 ± 419 93 ± 4 5±1 7,099 ± 452 5,349 ± 521 74 ± 3 94 ± 19 22.9 ± 0.2 0.23 ± 0.02 Normoxia 322 ± 6 76 ± 5* 39 ± 3** 5±1 2,594 ± 413 1,978 ± 370 87 ± 1 3±1 4,351 ± 196* 2,523 ± 311* 72 ± 3 310 ± 51* 19.1 ± 0.8* 0.32 ± 0.01* Hypoxia 430 ± 58 154 ± 15 25 ± 2 3±1 1,509 ± 153 1,040 ± 133 86 ± 5 4±1 4,747 ± 330 3,699 ± 330 71 ± 6 84 ±12 20.1 ± 0.7 0.23 ± 0.02 330 ± 19 144 ± 16 28 ± 2 4±1 1,765 ± 174 1,248 ± 134 92 ± 4 6±1 4,918 ± 171 3,659 ± 319 76 ± 2 97 ± 8 19.3 ± 0.6 0.26 ± 0.02 Data represent mean ± SEM. *, P < 0.05; **, P < 0.01, normoxia versus hypoxia, as determined by Kruskal-Wallis analysis of variance followed by Dunn’s multiple comparison test. RV/LV&septum, right ventricle/left ventricle including septum. Table S4. Hemodynamic data from ApoE/ mouse procedures Phenotype parameter Chow Heart rate (BPM) Cardiac index (µl/min/g) RVSP (mm Hg) RVEDP (mm Hg) RV dP/dtmax (mm Hg/s) RV dP/dtmin (mm Hg/s) LVESP (mm Hg) LVEDP (mm Hg) LV dP/dtmax (mm Hg/s) LV dP/dtmin (mm Hg/s) Mean Aortic pressure (mm Hg) ePVRi (mm Hg/ml/min/g) PA AT (ms) RV/LV&Septum ApoE/ TRAIL/ Paigen 337 ± 17 375 ± 11 155 ± 16 139 ± 16 25 ± 1 23 ± 2 2±1 2±1 1,325 ± 192 1,720 ± 180 1,140 ± 178 1,405 ± 117 85 ± 6 87 ± 3 4±1 4±1 7,873 ± 818 7,466 ± 1010 6,934 ± 789 6,764 ± 983 69 ± 7 68 ± 5 90 ± 12 91 ± 17 20.7 ± 1.4 19.7 ± 0.2 0.20 ± 0.01 0.22 ± 0.02 Chow 413 ± 19 128 ± 19 26 ± 2 2±1 1,927 ± 145 2,107 ± 776 87 ± 3 4±1 6,121 ± 436 4,959 ± 564 62 ± 10 119 ±16 19.8 ± 0.1 0.21 ± 0.01 ApoE//TRAIL/ Paigen Chow Paigen 385 ± 17 95 ± 18 50 ± 4*** 7 ± 1*** 3,092 ± 248 2,940 ± 467 86 ± 2 5±1 8,475 ± 843 7,523 ± 853 67 ± 2 318 ± 36*** 17.3 ± 0.2* 0.24 ± 0.01 428 ± 24 133 ± 14 26 ± 1 1±1 2,144 ± 300 1,457 ± 260 87 ± 4 3±1 8,005 ± 1260 4,565 ± 1144 65 ± 5 113 ± 9 20.7 ± 1.3 0.21 ± 0.01 393 ± 28 151 ± 30 29 ± 4### 1 ± 2### 2,358 ± 246 2,351 ± 1094 85 ± 3 4±1 4,815 ± 321 3,741 ± 587 63 ± 6 108 ± 7### 19.8 ± 1.2 0.22± 0.01 Data represent mean ± SEM. *, P < 0.05; ***, P < 0.001, chow versus Paigen diet. ###, P < 0.001, ApoE/ /TRAIL/ versus ApoE/ as determined by one-way ANOVA followed by Bonferroni’s multiple comparison test. RV/LV&septum, right ventricle/left ventricle including septum. S2 TRAIL drives PAH pathogenesis | Hameed et al. Downloaded from jem.rupress.org on October 22, 2012 Heart rate (BPM) Cardiac index (µl/min/g) RVSP (mm Hg) RVEDP (mm Hg) RV dP/dtmax (mm Hg/s) RV dP/dtmin (mm Hg/s) LVESP (mm Hg) LVEDP (mm Hg) LV dP/dtmax (mm Hg/s) LV dP/dtmin (mm Hg/s) Mean aortic pressure (mm Hg) ePVRi (mm Hg/ml/min/g) PA AT (ms) RV/LV&Septum TRAIL/ Published October 15, 2012 Table S5. Hemodynamic data from ApoE/ mouse intervention studies Phenotype parameter ApoE//TRAIL/ Saline rTRAIL 415 ± 22 273 ± 71 23 ± 2 3±1 2,674 ± 549 1,932 ± 741 94 ± 1 5±1 8,124 ± 1350 5,343 ± 931 79 ± 6 133 ± 27 19.8 ± 1.5 0.20 ± 0.01 +IgG 387 ± 66 193 ± 32 52 ± 1* 5±2 3,742 ± 815 3,006 ± 454 87 ± 4 3±1 7,642 ± 406 5,149 ± 276 71 ± 5 471 ± 88** 17.7 ± 1 0.34 ± 0.02 +TRAIL Ab 362 ± 28 161± 22 88 ± 18 6±1 7,616 ± 2227 3,516± 528 86 ± 4 4±1 6,177 ± 567 5,442 ± 379 73 ± 3 484 ±116 17.9 ± 1.2 0.22 ± 0.01 414 ± 17 223 ± 12 30 ± 3# 2 ± 1## 2,009 ± 208 1,355 ± 336 97 ± 1 5±1 7,540 ± 325 5,938 ± 369 75 ± 4 102 ± 12## 19.3 ± 0.3 0.21 ± 0.01 Data represent mean ± SEM. *, P < 0.05; **, P < 0.01, rTRAIL versus saline. #, P < 0.05; ##, P < 0.01, anti-TRAIL antibody versus IgG as determined by Mann-Whitney U test. RV/ LV&septum, right ventricle/left ventricle including septum. Table S6. Hemodynamic data from BMT mouse procedures Phenotype parameter BM donor/BM recipient ApoE/ into ApoE/ Chow Heart rate (BPM) Cardiac index (µl/min/g) RVSP (mm Hg) RVEDP (mm Hg) RV dP/dtmax (mm Hg/s) RV dP/dtmin (mm Hg/s) Paigen 385 ± 8 353 ± 18 153 ± 21 112 ± 7 26 ± 1 36 ± 5* 3±1 3±1 2,339 ± 363 3,415 ± 935 1,430 ± 1,792 ± 188 344 LVESP (mm Hg) 91 ± 1 84 ± 5 LVEDP (mm Hg) 4±1 4±1 LV dP/dtmax (mm Hg/s) 6,603 ± 1104 6,839 ± 2539 LV dP/dtmin (mm Hg/s) 4,900 ± 4,370 ± 997 871 Mean aortic pressure (mm Hg) 76 ± 7 76 ± 5 ePVRi (mm Hg/ml/min/g) 147 ± 23 282 ± 81* PA AT (ms) 22.3 ± 1.9 16.7 ± 1.0 0.23 ± 0.01 0.26 ± 0.02 RV/LV&Septum ApoE/ into ApoE//TRAIL/ ApoE//TRAIL/ into ApoE/ Chow Paigen Chow 435 ± 30 167 ± 17 23 ± 1 2±1 1,954 ± 125 375 ± 24 138 ± 20 30 ± 1 4±1 2,442 ± 94 375 ± 15 158 ± 19 26 ± 3 2±2 2,290 ± 486 Paigen ApoE//TRAIL/ into ApoE//TRAIL/ Chow Paigen 1,779 ± 1,919 ± 263 326 93 ± 2 87 ± 3 4±1 5±1 6,147 ± 582 6,270 ± 1136 382 ± 9 403 ± 35 385 ± 31 114 ± 10 146 ± 8 162 ± 27 47 ± 6** 25 ± 1 24 ± 3 5±1 2±1 2±1 5,701 ± 1,829 ± 417 1,908 ± 132 2623 1,301 ± 28 5,608 ± 1,702 ± 121 1,303 ± 157 2263 82 ± 2 82 ± 2 76 ± 4 81 ± 8 3±1 3±1 4±1 3±1 5,802 ± 476 4,802 ± 309 5,703 ± 1252 5,358 ± 1203 5,310 ± 3,876 ± 230 550 74 ± 4 70 ± 1 116 ±15 199 ± 34 24.0 ± 0.1 22.2 ± 0.6 0.26 ± 0.01 0.27 ± 0.01 4,542 ± 487 3,955 ± 365 65 ± 4 68 ± 2 154 ± 49 396 ± 89* 19.7 ± 1.5 15.2 ± 0.6 0.23± 0.01 0.28 ± 0.01 4,759 ± 443 3,313 ± 527 74 ± 5 147 ± 12 22.7 ± 2.7 0.23± 0.01 68 ± 2 133 ± 43 24.3 ± 3.1 0.23 ± 0.01 Data represent mean ± SEM. *, P < 0.05; **, P < 0.01, chow versus Paigen diet as determined by one-way ANOVA followed by Bonferroni’s multiple comparison test. RV/ LV&septum, right ventricle/left ventricle including septum. JEM View publication stats S3 Downloaded from jem.rupress.org on October 22, 2012 Heart rate (BPM) Cardiac index (µl/min/g) RVSP (mm Hg) RVEDP (mm Hg) RV dP/dtmax (mm Hg/s) RV dP/dtmin (mm Hg/s) LVESP (mm Hg) LVEDP (mm Hg) LV dP/dtmax (mm Hg/s) LV dP/dtmin (mm Hg/s) Mean aortic pressure (mm Hg) ePVRi (mm Hg/ml/min/g) PA AT (ms) RV/LV&Septum ApoE/ Paigen