Supplementary Materials

Supplementary Material for:

Blocking Macrophage Leukotriene B4 Prevents Endothelial Injury and Reverses Pulmonary Hypertension

Wen Tian, Xinguo Jiang, Rasa Tamosiuniene, Yon K. Sung, Jin Qian, Gundeep Dhillon, Lajos Gera, Laszlo Farkas, Marlene Rabinovitch, Roham T. Zamanian, Mohammed Inayathullah, Marina Fridlib, Jayakumar Rajadas, Marc Peters-Golden, Norbert F. Voelkel, Mark R. Nicolls*

*Corresponding author. E-mail: mnicolls@stanford.edu

Published 28 August 2013, Sci. Transl. Med. 5, 200ra117 (2013)
DOI: 10.1126/scitranslmed.3006674

This PDF file includes:

  • Materials and Methods
  • Fig. S1. Increased macrophage 5-LO during the evolution of experimental PH.
  • Fig. S2. Tissue-specific 5-LO expression in experimental PH.
  • Fig. S3. Increased nuclear membrane–translocated p5-LO over time as an indicator for 5-LO activation.
  • Fig. S4. High concentration of iNOS+ macrophages around occluded arterioles.
  • Fig. S5. In vivo NO release decreased in lungs from PH rats.
  • Fig. S6. Cellular ROS production is elevated in the SU group.
  • Fig. S7. Prevention of PH by macrophage depletion.
  • Fig. S8. Induction of PAEC apoptosis by interstitial lung macrophage (IMØ)– derived LTB4.
  • Fig. S9. Experimental groups of PAEC-macrophage coculture system.
    Fig. S10. Induction of PAEC apoptosis by alveolar macrophage (AMØ)–derived LTB4.
  • Fig. S11. LTB4 induction of PAEC apoptosis in a dose-dependent manner.
  • Fig. S12. LTB4 induction of synchronized PAEC apoptosis in a dose-dependent manner.
  • Fig. S13. S1P rescues PAEC apoptosis induced by macrophage LTB4.
  • Fig. S14. Quantification of Western blots evaluating LTB4-mediated PAEC apoptosis.
  • Fig. S15. The reversal of established PH by administration of bestatin 3 weeks after SU administration.
  • Fig. S16. The improvement of right heart function of PH animals undergoing blockade of LTB4 signaling.
  • Fig. S17. The dose-dependent reversal of established PH using novel oral formulations of bestatin.
  • Fig. S18. The reversal of established PH using a novel formulation of inhaled bestatin.
  • Fig. S19. Blocking multiple eicosanoid pathways in the SU/athymic rat PH model.
  • Fig. S20. Reduction of disease-related LTA4H expression in macrophages in bestatin-treated rats.
  • Fig. S21. Inflammation attenuation in bestatin-treated animals.
  • Fig. S22. Prevention of PH lung macrophage–induced PAEC apoptosis with bestatin.
  • Fig. S23. Reestablishing lung Sphk1-eNOS signaling with bestatin treatment in established PH.
  • Fig. S24. Prevention of mutant-mimic macrophage-induced PAEC apoptosis and maintenance of Sphk1-eNOS signaling with exogenous bestatin.
  • Fig. S25. The induction of cellular apoptosis in remodeled PH vessel wall 1 week after bestatin treatment.
  • Fig. S26. The reversal of PH in the MCT model using bestatin therapy.
  • Fig. S27. The failure of bestatin in the low-LTB4 SU/hypoxia PH model.
  • Table S1. Hemodynamic and echocardiographic data for athymic rats at different time points after SU administration.
  • Table S2. Dosing regimen for different eicosanoid inhibitors.

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