The aberrant expression and localization of DNA methyltransferase 3B in endometriotic stromal cells Matthew T. Dyson, Ph.D. * , Toshiyuki Kakinuma, M.D. * , Mary Ellen Pavone, M.D., Diana Monsivais, Ph.D., Antonia Navarro, Ph.D., Saurabh S. Malpani, M.S., Masanori Ono, M.D., and Serdar E. Bulun, M.D. Division of Reproductive Biology Research, Northwestern University Feinberg School of Medicine, Chicago, IL Abstract Objective—To define the expression and function of DNA methyltransferases (DNMTs) in response to decidualizing stimuli in endometriotic cells compared with healthy endometrial stroma. Design—Basic science. Setting—University research center. Patients—Premenopausal women with or without endometriosis. Interventions—Primary cultures of stromal cells from healthy endometrium (E-IUM) or endometriomas (E-OSIS) were subjected to in vitro decidualization (IVD) using 1 µM medroxyprogesterone acetate, 35 nM 17β-estradiol, and 0.05 mM 8-Br-cAMP. Main Outcome Measure(s)—DNMT1, DNMT3A, and DNMT3B expression in E-IUM and E- OSIS were assessed by qRT-PCR and immunoblotting. DNMT3B recruitment to the promoters of steroidogenic factor 1 (SF-1) and estrogen receptor α (ESR1) was examined by chromatin immunoprecipitation Results—IVD treatment reduced DNMT3B mRNA (74%) and protein levels (81%) only in E- IUM. DNMT1 and DNMT3A were unchanged in both cell types. Significantly more DNMT3B bound to the SF-1 promoter in E-IUM compared with E-OSIS, and IVD treatment reduced binding in E-IUM to levels similar to those in E-OSIS. DNMT3B enrichment across three ESR1 promoters was reduced in E-IUM after IVD, although the more distal promoter showed increased DNMT3B enrichment in E-OSIS after IVD. Corresponding Author: Matthew T. Dyson., Robert H. Lurie Medical Research Center, Room 4-123, 303 E. Superior Chicago, IL 60611, m-dyson@northwestern.edu, 312-503-5281 (tel), 312-503-0095 (fax). * These authors should be considered similar in author order. Publisher's Disclaimer: This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final citable form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. Disclosure Statement: M.T.D. has nothing to disclose. T.K. has nothing to disclose. M.E.P. has nothing to disclose. D.M. has nothing to disclose. A.N. has nothing to disclose. S.S.M. has nothing to disclose. M.O. has nothing to disclose. S.E.B. has nothing to disclose. HHS Public Access Author manuscript Fertil Steril. Author manuscript; available in PMC 2016 October 01. Published in final edited form as: Fertil Steril. 2015 October ; 104(4): 953–963.e2. doi:10.1016/j.fertnstert.2015.06.046. Author Manuscript Author Manuscript Author Manuscript Author Manuscript