Expression of the novel gene embryo implantation factor 2 (EMO2) in the mouse uterus at the implantation sites Zhaogui Sun, Ph.D., Renwei Su, Ph.D.,

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Expression of the novel gene embryo implantation factor 2 (EMO2) in the mouse uterus at the implantation sites Zhaogui Sun, Ph.D., Renwei Su, Ph.D., Zengming Yang, Ph.D., Huijuan Shi, Ph.D., Chengquan Liu, B.Sc., Jian Wang, Ph.D. Fertility and Sterility Volume 91, Issue 5, Pages 2116-2122 (May 2009) DOI: 10.1016/j.fertnstert.2008.01.092 Copyright © 2009 American Society for Reproductive Medicine Terms and Conditions

Figure 1 Northern blot analysis of EMO2 mRNA in the uterine tissues of day 5 implantation sites (D5I) and day 5 interimplantation sites (D5i-I). The lower panel shows the 18s RNA bands of each RNA sample. Fertility and Sterility 2009 91, 2116-2122DOI: (10.1016/j.fertnstert.2008.01.092) Copyright © 2009 American Society for Reproductive Medicine Terms and Conditions

Figure 2 The tissue distribution of EMO2 mRNA was analyzed by RT-PCR by using GAPDH mRNA as the control. EMO2 mRNA was detected in all of the tested tissues. B = brain; Lu = lung; H = heart; Li = liver; Sp = spleen; K = kidney; M = muscle; SI = small intestine; Sk = skin; T = testis; E = epididymis; O = ovary; U = uterus. Fertility and Sterility 2009 91, 2116-2122DOI: (10.1016/j.fertnstert.2008.01.092) Copyright © 2009 American Society for Reproductive Medicine Terms and Conditions

Figure 3 The antigen specificity of rabbit anti-EMO2 antisera was examined by Western blot analysis. Bacteria GST-EMO2/BL21 lysate [1], bacteria BL21 lysate [2], and bacteria GST/BL21 lysate [3] were subjected to SDS-PAGE. The anti-GST antibody (A), rabbit anti-EMO2 sera (B), and normal rabbit sera (C) were, respectively, used as the primary antibodies. The prepared rabbit anti-EMO2 antibody specifically recognized the fusion protein GST-EMO2 (∼53 kDa) that is contained in bacteria GST-EMO2/BL21 lysate but did not bind the GST protein (∼26 kDa) that is contained in bacteria GST/BL21 lysate. Fertility and Sterility 2009 91, 2116-2122DOI: (10.1016/j.fertnstert.2008.01.092) Copyright © 2009 American Society for Reproductive Medicine Terms and Conditions

Figure 4 Immunohistochemical staining of mouse uterus for the EMO2 protein using rabbit anti-EMO2 serum as the primary antibody. The uterine sections were stained with EMO2 for day 1 pregnant uterus (A), day 4 pregnant uterus (B), implantation sites (C), and interimplantation sites (D) on day 5 of pregnancy, implantation sites on days 6 (E), 7 (F), and 8 (G) of pregnancy, day 4 pseudopregnant uterus (I), day 5 pseudopregnant uterus (J), delayed implantation before (K), and 24 hours after (L) the activation, artificially induced decidualization (AID) of pseudopregnant uterus (M) and the control horn of AID (N), P-treated uterus (O), and oil-treated uterus (P). Panel H is the negative control for panel G, in which preimmunized normal rabbit serum is substituted for the primary antibody. l = lumen of uterus; g = gland; sc = stromal cells; dc = decidual cells; em = embryo. Scale bars represents 10 μm. Fertility and Sterility 2009 91, 2116-2122DOI: (10.1016/j.fertnstert.2008.01.092) Copyright © 2009 American Society for Reproductive Medicine Terms and Conditions