Aimee S. Browne, M. D. , M. Sc. , Jie Yu, M. D. , M. Sc

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Proteomic identification of neurotrophins in the eutopic endometrium of women with endometriosis Aimee S. Browne, M.D., M.Sc., Jie Yu, M.D., M.Sc., Ruo-Pan Huang, M.D., Ph.D., Antônio M.C. Francisco, M.D., Neil Sidell, Ph.D., Robert N. Taylor, M.D., Ph.D. Fertility and Sterility Volume 98, Issue 3, Pages 713-719 (September 2012) DOI: 10.1016/j.fertnstert.2012.05.027 Copyright © 2012 American Society for Reproductive Medicine Terms and Conditions

Figure 1 (A) Reverse transcriptase–polymerase chain reaction confirmed the expression of NGF mRNA (predicted amplicon = 340 bp) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mRNA (predicted amplicon = 185 bp) in eutopic endometrial biopsies from two independent and representative controls (C) and two independent endometriosis (E) subjects. Amplicon lengths are indicated as base pairs (bp) and correspond with the molecular weight (MW) ladder in the left lane. (B) Reverse transcriptase–polymerase chain reaction confirmed the expression of BDNF mRNA (predicted amplicon = 120 bp), NT-4/5 mRNA (predicted amplicon = 96 bp), and GAPDH mRNA (predicted amplicon = 185 bp) in eutopic endometrial biopsies from a representative control (C) and two independent endometriosis (E) subjects. GAPDH amplification was performed in duplicate. Molecular weight (MW) ladder is shown at left. Fertility and Sterility 2012 98, 713-719DOI: (10.1016/j.fertnstert.2012.05.027) Copyright © 2012 American Society for Reproductive Medicine Terms and Conditions

Figure 2 (A) Quantitative ELISAs of NGF and NT-4/5 proteins in lysates of endometrial biopsies from endometriosis (n = 18) and control (n = 15) subjects. Data are shown as mean ± SD (*P=.04, Mann-Whitney U test). (B) Quantitative ELISA of BDNF protein in lysates of endometrial biopsies from endometriosis (n = 18) and control (n = 15) subjects. Data are shown as mean ± SD (**P<.01, Mann-Whitney U test). Fertility and Sterility 2012 98, 713-719DOI: (10.1016/j.fertnstert.2012.05.027) Copyright © 2012 American Society for Reproductive Medicine Terms and Conditions

Figure 3 (A) Western blot of NGF in lysates of endometrial biopsies. Two representative controls (C) and endometriosis (E) cases are shown. Molecular weights are shown in kilodaltons as determined from the standard ladder shown at the right (MW). The faint band at 13 kD represents mature NGF processed from the 27 kD precursor. β-actin, a housekeeping control protein, was immunoblotted to verify equal loading. (B) Western blots of NT-4/5 and BDNF in lysates of endometrial biopsies. Two representative controls (C) and endometriosis (E) cases are shown. Molecular weights are shown in kilodaltons at the right. The predominant bands of 22 kD and 28 kD represent proNT-4/5 and proBDNF, respectively. The faint bands at 14 kD represent mature, processed NT-4/5 and BDNF. β-actin, a housekeeping control protein, was immunoblotted to verify equal loading. Fertility and Sterility 2012 98, 713-719DOI: (10.1016/j.fertnstert.2012.05.027) Copyright © 2012 American Society for Reproductive Medicine Terms and Conditions