Panagiotis Bakas, M. D. , Angelos Liapis, M. D

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Estrogen receptor α and β in uterine fibroids: a basis for altered estrogen responsiveness Panagiotis Bakas, M.D., Angelos Liapis, M.D., Spiros Vlahopoulos, Ph.D., Maria Giner, M.D., Stella Logotheti, B.Sc., Georgios Creatsas, M.D., Aggeliki K. Meligova, B.Sc., Michael N. Alexis, Ph.D., Vassilis Zoumpourlis, Ph.D. Fertility and Sterility Volume 90, Issue 5, Pages 1878-1885 (November 2008) DOI: 10.1016/j.fertnstert.2007.09.019 Copyright © 2008 American Society for Reproductive Medicine Terms and Conditions

Figure 1 Estrogen receptor levels (median values) in the cytosol of leiomyomas (LEIO) compared with normal myometrium (MYOM) from 35 patients operated on for symptomatic fibroids. Median values of ER levels in leiomyoma and the corresponding myometrium samples, as assessed by Scatchard plot analysis with use of tritiated E2, were calculated by using the nonparametric Wilcoxon paired samples test, expressed as femtomoles of ER per milligram of cytosol protein and plotted with use of a box-and-whisker graph. The boxes contain the values between the upper (75th percentile) and lower (25th percentile) quartiles, the lines across the boxes correspond to the medians, and the whiskers extend to the highest and lowest values, excluding outliers (open square) and extreme values (filled squares). ∗Significantly different compared with myometrium (P<.05). Fertility and Sterility 2008 90, 1878-1885DOI: (10.1016/j.fertnstert.2007.09.019) Copyright © 2008 American Society for Reproductive Medicine Terms and Conditions

Figure 2 Estrogen receptor α and ERβ mRNA levels in leiomyomas and the corresponding normal myometrium, as compared with the respective GAPDH mRNA levels. Estrogen receptor α, ERβ, and GAPDH mRNA levels in the myometrium (M) and the corresponding leiomyoma samples (L) from each individual patient were obtained by semiquantitative multiplex RT-PCR. Typical ethidium bromide–stained gels of the RT-PCR reaction products from the samples of 20 patients are shown. Fertility and Sterility 2008 90, 1878-1885DOI: (10.1016/j.fertnstert.2007.09.019) Copyright © 2008 American Society for Reproductive Medicine Terms and Conditions

Figure 3 Protein levels and DNA-binding status of ERα in leiomyomas and the corresponding normal myometrium. (A) Representative immunoprecipitation, PAGE, and immunoblotting analysis of ERα in nuclear extracts from normal myometrium (M) and leiomyoma (L) samples no. 5 (5M and 5L) and no. 10 (10M and 10L), selected for their high ERα-to-ERβ mRNA expression ratio (see samples numbered in Fig. 2). (B) vitERE binding of protein in the nuclear extracts from normal myometrium and leiomyoma samples no. 5 and no. 10, as assessed by EMSA (lanes 1 vs. 2 and 3 vs. 4, respectively). Supershifting of vitERE-binding of protein in the nuclear extracts from normal myometrium and leiomyoma sample no. 10, as assessed by EMSA in the presence of antibodies to ERα (a-ERα, lanes 5 and 8) and ERβ (a-ERβ, lanes 6 and 7). ∗Significantly different compared with 5M (P<.05; t-test). #Significantly different compared with 10M (P<.05; t-test). Ab = antibodies. Fertility and Sterility 2008 90, 1878-1885DOI: (10.1016/j.fertnstert.2007.09.019) Copyright © 2008 American Society for Reproductive Medicine Terms and Conditions