Marina Ines Flamini, Ph. D. , Angel Matias Sanchez, Ph. D

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Estrogen regulates endometrial cell cytoskeletal remodeling and motility via focal adhesion kinase Marina Ines Flamini, Ph.D., Angel Matias Sanchez, Ph.D., Andrea Riccardo Genazzani, M.D., Ph.D., Tommaso Simoncini, M.D., Ph.D. Fertility and Sterility Volume 95, Issue 2, Pages 722-726 (February 2011) DOI: 10.1016/j.fertnstert.2010.08.039 Copyright © 2011 American Society for Reproductive Medicine Terms and Conditions

Figure 1 (A) Ishikawa cell treatment with E2 (10−10–10−7 M) and with E2 (10−9 M) in the presence of ICI 182,780 or (B) with E2 (10−9 M) for 5, 10, 15, 30, or 60 minutes. p-FAK Tyr397, p-FAK Tyr576, FAK, and actin are shown. ∗P≤.05 vs. control. Fertility and Sterility 2011 95, 722-726DOI: (10.1016/j.fertnstert.2010.08.039) Copyright © 2011 American Society for Reproductive Medicine Terms and Conditions

Figure 2 Ishikawa cell treatment with E2 in the presence or absence of ICI 182,780 (ICI) and (A) Texas Red-phalloidin linked to actin fibers, p-FAK Tyr397 linked to fluorescein isothiocyanate conjugate, (B) vinculin fibers linked to Texas Red, and p-FAK Tyr397 linked to fluorescein isothiocyanate conjugate. Yellow arrows indicate membrane ruffles and pseudopodia. Fertility and Sterility 2011 95, 722-726DOI: (10.1016/j.fertnstert.2010.08.039) Copyright © 2011 American Society for Reproductive Medicine Terms and Conditions

Figure 3 (A) Ishikawa cell treatment with E2, PPT, or DPN in the presence or absence of ICI 182,780 (ICI). (B) Ishikawa cell treatment with E2 in the presence or absence of different inhibitors. (C) Ishikawa cell treatment with E2 in the presence or absence of ICI 182,780. Protein extracts were immunoprecipitated with an antibody against ERα, and the IPs were assayed for coimmunoprecipitation of FAK, p85α, and c-Src. (D) Ishikawa cell treatment with E2 in the presence or absence of ICI 182,780. Protein extracts were immunoprecipitated with an antibody against ERβ, and the IPs were assayed for coimmunoprecipitation of FAK, p85α, and c-Src. IP = immunoprecipitate. Fertility and Sterility 2011 95, 722-726DOI: (10.1016/j.fertnstert.2010.08.039) Copyright © 2011 American Society for Reproductive Medicine Terms and Conditions

Figure 4 Treatment of ESC with E2 (48 hours) in the presence or absence of WM or PTX or transfect with FAK siRNA. (A) Horizontal migration images. (B) Mean migration length quantitative analysis. (C) Matrigel chamber images. (D) Mean number of invading cells. Data are expressed as mean ± SD of three separate experiments. ∗∗P≤.05 vs. control; ∗P≤.05 vs. E2. Fertility and Sterility 2011 95, 722-726DOI: (10.1016/j.fertnstert.2010.08.039) Copyright © 2011 American Society for Reproductive Medicine Terms and Conditions