Glycodelin A is expressed differentially in normal human endometrial tissue throughout the menstrual cycle as assessed by immunohistochemistry and in situ hybridization  Ioannis Mylonas, M.D., Udo Jeschke, Ph.D., Christiane Kunert-Keil, Ph.D., Naim Shabani, M.D., Darius Dian, M.D., Ingo Bauerfeind, M.D., Christina Kuhn, Markus S. Kupka, M.D., Ph.D., Klaus Friese, M.D., Ph.D.  Fertility and Sterility  Volume 86, Issue 5, Pages 1488-1497 (November 2006) DOI: 10.1016/j.fertnstert.2006.03.062 Copyright © 2006 American Society for Reproductive Medicine Terms and Conditions

FIGURE 1 Immunohistochemical expression of glycodelin. Whereas the monoclonal antibody showed no or minimal expression during the proliferative phase (A), the polyclonal-peptide antibody produced a clear, albeit weak, staining reaction (B). During the early secretory phase, the staining reaction with the monoclonal antibody was confined to glandular epithelial cells (C), and a more intense staining reaction was observed for the polyclonal-peptide antibody (D). The most intense reaction was observed during the late secretory phase for the monoclonal antibody (E), whereas the polyclonal-peptide antibody reacted also with the strongest intensity during this menstrual phase (F). Magnification, all panels, ×250. Mylonas. Glycodelin expression in human endometrium. Fertil Steril 2006. Fertility and Sterility 2006 86, 1488-1497DOI: (10.1016/j.fertnstert.2006.03.062) Copyright © 2006 American Society for Reproductive Medicine Terms and Conditions

FIGURE 2 Evaluation of immunohistochemical glycodelin staining reaction. (A) The IRS showed a significant rise between the proliferative and late secretory phases (*P<.002) for the glycodelin monoclonal antibody, whereas the polyclonal glycodelin peptide antibody showed a similar rise but without statistical significance. (B) The staining intensity demonstrated a significant rise between the proliferative and early secretory (*P<.05) as well as between the proliferative and late secretory phases (**P<.05). All data are means ± SEM. Significance was assumed at P<.05 (all asterisked values). Mylonas. Glycodelin expression in human endometrium. Fertil Steril 2006. Fertility and Sterility 2006 86, 1488-1497DOI: (10.1016/j.fertnstert.2006.03.062) Copyright © 2006 American Society for Reproductive Medicine Terms and Conditions

FIGURE 3 Glycodelin mRNA was expressed minimally in endometrial tissue from the proliferative phase (A, antisense; B, sense). Glycodelin mRNA was expressed minimally in endometrial tissue from the proliferative phase (C, antisense; D, sense). Glycodelin mRNA decreased during the late secretory phase (E, antisense; F, sense). Magnification, all panels, ×100. Mylonas. Glycodelin expression in human endometrium. Fertil Steril 2006. Fertility and Sterility 2006 86, 1488-1497DOI: (10.1016/j.fertnstert.2006.03.062) Copyright © 2006 American Society for Reproductive Medicine Terms and Conditions

FIGURE 4 Evaluation of glycodelin expression by in situ hybridization. Staining intensity of glycodelin mRNA by in situ hybridization reaction in endometrial tissue as determined by computerized analysis showed a significant rise of mRNA expression during the early secretory phase compared with during the proliferative phase (*P<.05). Data are means ± SEM. Significance was assumed at P<.05 (asterisked values). Mylonas. Glycodelin expression in human endometrium. Fertil Steril 2006. Fertility and Sterility 2006 86, 1488-1497DOI: (10.1016/j.fertnstert.2006.03.062) Copyright © 2006 American Society for Reproductive Medicine Terms and Conditions

FIGURE 5 Binding of glycodelin monoclonal and polyclonal peptide antibodies to deglycosylated endometrial tissue. Both antibodies reacted positively primarily with endometrial glandular epithelial cells from the late secretory phase (A and C). After the deglycosylation procedure, the staining reaction of the monoclonal antibody was minimal within the glandular epithelial cells, showing a substantial decrease in the intensity (B). The immunoreaction of the polyclonal-peptide antibody was localized in epithelial cells, with a diffuse staining reaction within the endometrial stroma (C). We found no significant decrease of the staining reaction after the deglycosylation process (D). Magnification, all panels, ×250. Mylonas. Glycodelin expression in human endometrium. Fertil Steril 2006. Fertility and Sterility 2006 86, 1488-1497DOI: (10.1016/j.fertnstert.2006.03.062) Copyright © 2006 American Society for Reproductive Medicine Terms and Conditions