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Kocanova et al. BMC Cell Biology 2010, 11:98 /1471-2121/11/98 RESEARCH ARTICLE Open Access Ligands specify estrogen receptor alpha nuclear localization and degradation Silvia Kocanova1,2†, Mahta Mazaheri1,2†, Stéphanie Caze-Subra1,2, Kerstin Bystricky1,2* Abstract Background: The estrogen receptor alpha (ERa) is found predominately in the nucleus, both in hormone stimulated and untreated cells. Intracellular distribution of the ERa changes in the presence of agonists but the impact of different antiestrogens on the fate of ERa is a matter of debate. Results: A MCF-7 cell line stably expressing GFP-tagged human ERa (SK19 cell line) was created to examine the localization of ligand-bound GFP-ERa. We combined digitonin-based cell fractionation analyses with fluorescence and immuno-electron microscopy to determine the intracellular distribution of ligand-bound ERa and/or GFP-ERa. Using fluorescence- and electron microscopy we demonstrate that both endogenous ERa and GFP-ERa form numerous nuclear focal accumulations upon addition of agonist, 17b-estradiol (E2), and pure antagonists (selective estrogen regulator disruptor; SERD), ICI 182,780 or RU58,668, while in the presence of partial antagonists (selective estrogen regulator modulator; SERM), 4-hydroxytamoxifen (OHT) or RU39,411, diffuse nuclear staining persisted. Digitonin based cell fractionation analyses confirmed that endogenous ERa and GFP-ERa predominantly reside in the nuclear fraction. Overall ERa protein levels were reduced after estradiol treatment. In the presence of SERMs ERa was stabilized in the nuclear soluble fraction, while in the presence of SERDs protein levels decreased drasti- cally and the remaining ERa was largely found in a nuclear insoluble fraction. mRNA levels of ESR1 were reduced compared to untreated cells in the presence of all