10.6084/m9.figshare.4263686.v1 Chen T. Chen T. Chen N. Chen N. Pang N. Pang N. Xiao L. Xiao L. Li Y. Li Y. Li R. Li R. Luo M. Luo M. Deng X. Deng X. Ren M. Ren M. Wu J. Wu J. Wang L. Wang L. Supplementary Material for: Hydroxysafflor Yellow A Promotes Angiogenesis via the Angiopoietin 1/ Tie-2 Signaling Pathway Karger Publishers 2016 Hydroxysafflor yellow A Angiogenesis Angiopoietin 1 Tie-2 Hindlimb ischemia 2016-11-28 15:11:17 Figure https://karger.figshare.com/articles/figure/Supplementary_Material_for_Hydroxysafflor_Yellow_A_Promotes_Angiogenesis_via_the_Angiopoietin_1_Tie-2_Signaling_Pathway/4263686 <i>Background:</i> The flowers of <i>Carthamus tinctorius</i> L. are widely used in traditional Chinese medicine to treat cerebrovascular and cardiovascular diseases. Hydroxysafflor yellow A (HSYA), the main constituent of <i>C. tinctorius</i> L. flowers, is known for its multiple biological activities. The present study investigated the effects of HSYA on angiogenesis in vitro and in a mouse hindlimb ischemia model. <i>Methods:</i>Using human umbilical vein endothelial cells (HUVEC) in vitro and a mouse hindlimb ischemia model in vivo, the angiogenic role of HSYA was evaluated. <i>Results:</i> HSYA significantly increased the capillary-like tube formation and migration of HUVEC. HSYA not only induced a rise in the expression of angiopoietin 1 and Tie-2 but it also increased phosphorylation of Tie-2, Akt, and extracellular signal-regulated kinase 1/2. Furthermore, an anti-Tie-2 neutralizing antibody significantly inhibited HSYA-induced HUVEC tube formation and migration. In vivo, the recovery of perfusion of ischemic hindlimb tissue after femoral artery interruption was significantly increased in HSYA-treated mice compared to vehicle controls. Consistent with these results, the arteriole and capillary densities in ischemic gastrocnemius muscles were significantly increased in HSYA-treated mice. <i>Conclusions:</i> These results indicate the potential utility of HSYA for the treatment of ischemic diseases.