Maryam Hormozi, M. Sc. , Saeed Talebi, M. D. , Amir Hassan Zarnani, Ph

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5′-(N-ethylcarboxamido) adenosine improves angiogenesis in transplanted human ovarian tissue Maryam Hormozi, M.Sc., Saeed Talebi, M.D., Amir Hassan Zarnani, Ph.D., Mahmood Jeddi-Tehrani, Ph.D., Ladan Hosseini Gohari, Ph.D., Haleh Soltanghoraei, M.D., Mina Jafarabadi, M.D., Mohammad Mehdi Akhondi, Ph.D. Fertility and Sterility Volume 95, Issue 8, Pages 2560-2563.e5 (June 2011) DOI: 10.1016/j.fertnstert.2011.05.041 Copyright © 2011 American Society for Reproductive Medicine Terms and Conditions

Figure 1 5′-(N-ethylcarboxamido) adenosine (NECA) improves angiogenesis in transplanted human ovarian tissue. Nude mice were treated with NECA or vehicle after being transplanted with human ovarian tissues. At posttransplantation (PT) days 2, 7, and 30, ovarian fragments were recovered and the expression of (A) angiopoietin-1 (Ang1), (B) Ang2, (C) vascular endothelial growth factor 121 (VEGF-121), and (D) VEGF-189 in the NECA and control groups were assessed by quantitative real-time polymerase chain reaction. (E) Ang2:Ang1 ratio at different PT time intervals. All comparisons were made in reference to the pretransplantation state. Abs gene reg: absolute gene regulation. ∗P<.05, ∗∗P<.01. Fertility and Sterility 2011 95, 2560-2563.e5DOI: (10.1016/j.fertnstert.2011.05.041) Copyright © 2011 American Society for Reproductive Medicine Terms and Conditions

Supplemental Figure 1 Effect of 5′-(N-ethylcarboxamido) adenosine (NECA) on angiopoietin-1 (Ang1), Ang2, and vascular endothelial growth factor (VEGF) protein expression in heterotopic human ovarian xenografts. Nude mice were transplanted with human ovarian tissues and treated with NECA or vehicle as control. On posttransplantation (PT) days 2, 7, and 30, ovarian fragments were recovered, and the expression of Ang1, Ang2, and VEGF was analyzed by Western blotting using human-specific antibodies. Lane 1: Pretransplantation ovary. Lanes 2–4: PT days 2, 7, and 30 in the NECA group. Lanes 5–7: PT days 2, 7, and 30 the control group. Fertility and Sterility 2011 95, 2560-2563.e5DOI: (10.1016/j.fertnstert.2011.05.041) Copyright © 2011 American Society for Reproductive Medicine Terms and Conditions

Supplemental Figure 2 Effect of 5′-(N-ethylcarboxamido) adenosine (NECA) on vascular density of human ovarian xenograft. (A) Nude mice were treated with NECA (a–c) or vehicle (d–f) after being transplanted with human ovarian tissues. Ovarian fragments were recovered on days 2 (A-a, A-d), 7 (A-b, A-e), and 30 (A-c, A-f) after transplantation, and the vascular density was evaluated after hematoxylin and eosin staining. The vascular density of pretransplantation ovary is shown in A-g. Black arrows show blood vessels. (B) Quantitative analysis of vascular density in both groups. Fertility and Sterility 2011 95, 2560-2563.e5DOI: (10.1016/j.fertnstert.2011.05.041) Copyright © 2011 American Society for Reproductive Medicine Terms and Conditions