Gene and protein expression signature of endometrial glandular and stromal compartments during the window of implantation Gloria E. Evans, Ph.D., José.

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Gene and protein expression signature of endometrial glandular and stromal compartments during the window of implantation Gloria E. Evans, Ph.D., José A. Martínez-Conejero, Ph.D., Gregory T.M. Phillipson, M.B., Ch.B., Carlos Simón, M.D., Ph.D., Les A. McNoe, M.Sc., Peter H. Sykes, M.B., Ch.B., José A. Horcajadas, Ph.D., Enid Y.N. Lam, Ph.D., Cristin G. Print, Ph.D., Iris L. Sin, Ph.D., John J. Evans, Ph.D. Fertility and Sterility Volume 97, Issue 6, Pages 1365-1373.e2 (June 2012) DOI: 10.1016/j.fertnstert.2012.03.007 Copyright © 2012 American Society for Reproductive Medicine Terms and Conditions

Figure 1 (A) Top left: Messenger RNA (mRNA) expression from AFFYMETRIX testing of epithelial GLAND samples collected at LH+2 the nonreceptive phase (n = 9), all clustered quite distinctly from the receptive samples collected at LH+7 (n = 9, P=.05). (B) mRNA expression from AGILENT testing of epithelial GLAND samples collected at LH+2 the nonreceptive phase (n = 6), all clustered quite distinctly from the receptive samples collected at LH+7 (n = 5, P=.05). (C) mRNA expression from AFFYMETRIX testing of STROMAL samples collected at LH+2 the nonreceptive phase (n = 9), all clustered quite distinctly from the receptive samples collected at LH+7 (n = 9, P=.05). (D) The mRNA expression from AGILENT testing of STROMAL samples collected at LH+2 the nonreceptive phase (n = 6), all clustered quite distinctly from the receptive samples collected at LH+7 (n = 5, P=.05). Fertility and Sterility 2012 97, 1365-1373.e2DOI: (10.1016/j.fertnstert.2012.03.007) Copyright © 2012 American Society for Reproductive Medicine Terms and Conditions

Figure 2 Hinton plot. Independent component analysis (ICA) of messenger RNA (mRNA) expressed on AFFYMETRIX microarrays, 18 GLANDS and 18 STROMAL tissue compartments from nine fertile participants. This plot shows rows of the plot as independent components or mRNA signatures and columns as individual arrays. The area in each square is proportional to the magnitude of the contribution of each genechip to each component. Positive values are shown in yellow and negative values in red. The cluster analysis of glands at LH+2 and LH+7 are on two subbranches of the main left hand branch, while both groups of stroma cluster on the right-hand branch, although there are some outliers. Fertility and Sterility 2012 97, 1365-1373.e2DOI: (10.1016/j.fertnstert.2012.03.007) Copyright © 2012 American Society for Reproductive Medicine Terms and Conditions

Figure 3 (A) CCNB1 staining in green, showing nuclear staining in the stroma at LH+2, and (B) no staining at LH+7. (C) CCNB2 staining in red, showing staining in the nucleus, the nuclear pole, and the cytoplasm of the stroma with diffuse cytoplasmic staining in glands at LH+2 samples, and (D) staining only at the nuclear pole in the stroma and cytoplasmic staining in the glands at LH+7 sample. The cell nucleus is counterstained with DAPI. Fertility and Sterility 2012 97, 1365-1373.e2DOI: (10.1016/j.fertnstert.2012.03.007) Copyright © 2012 American Society for Reproductive Medicine Terms and Conditions

Supplemental Figure 1 Quantitative polymerase chain reaction. Line graphs comparing the relative values of messenger RNA (mRNA) expression between samples taken at LH+2 and LH+7 using SigmaStat. (A) CCNE2, (B) CCND1, (C) E2F3, and (D) SMAD3 mRNA expression in glands (purple) and (E) E2F2 mRNA expression in stroma (pink) between LH+2 and LH+7 from three of the nine participants in this sample set. P values were not calculated. Fertility and Sterility 2012 97, 1365-1373.e2DOI: (10.1016/j.fertnstert.2012.03.007) Copyright © 2012 American Society for Reproductive Medicine Terms and Conditions