SUPPORTINGINFORMATIONS.PDF

SUPPORTING INFORMATIONS Role of Sphingosine-1-phosphate in Transplant Vasculopathy evoked by anti-HLA antibody. 1,2 1,2 1 3 1 Magali Trayssac , Sylvain Galvani , Nathalie Augé , Roger Sabbadini , Denis Calise , Elodie Mucher 1,2 , Federico Sallusto 4 , Mogens Thomsen 1,5, Robert Salvayre 1,2, Anne Nègre-Salvayre 1 1 Inserm UMR-1048, Toulouse, France 2 University of Toulouse; Faculty of Medicine, Biochemistry Dept, Toulouse, France 3 LPath, Inc., and Department of Biology, San Diego State University, San Diego, CA 4 CHU Rangueil, Department of Nephrology, Toulouse, France 5 Inserm UMR-1027, Toulouse, France I – Materials Reagents Anti-human HLA class I antibody (clone W6/32, catalogue n° 857.220.000), anti-mouse major histocompatibility complex mAb (28-14-8, recognizing H-2Db), and isotype-matched non-specific control IgG2a mAb used to stimulate SMC were from Eurobio AbCys (Diaclone, Besançon, FR). As indicated by the manufacturer, the anti HLA antibody recognizes an antigenic determinant shared among products of the HLA-A, -B and -C loci. Anti human HLA Class I antibody (clone M-147) was from Abnova (Clinisciences, Nanterre, FR). Anti-SK1, anti-S1PR1 (ab23695) and anti-tubulin (ab44928) were from Abcam (Cambridge, UK), anti- S1PR3 antibody was from Imgenex (Novus Biological, Cliniscience, Nanterre, FR), anti-Na,K ATPase α1 subunit from Cell Signaling (Saint-Quentin-en-Yvelines, FR), anti-vimentin (clone SP20 rabbit mAb) from Epitomics (Burlingame, CA) and anti-α-smooth muscle actin (clone 1A4) and anti-β-actin from Sigma-Aldrich (Saint-Quentin Fallavier, FR), Alexa Fluor® 488 Goat Anti-Rabbit IgG Ab, Alexa Fluor® 488 Goat Anti-Mouse IgG Ab and Alexa Fluor® 546 Goat Anti-Rabbit IgG Ab f