Titlebook: Estrogen Receptors; Methods and Protocol Kathleen M. Eyster Book 2022Latest edition The Editor(s) (if applicable) and The Author(s), under

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Assessing Estrogenic Activity of Classical Estrogen Receptor -Binding Compounds, the estrogenic activity (i.e., both receptor-mediated agonism and/or antagonism) of these compounds. This chapter focuses on methods that allow determination of whether an ERα-binding compound acts as an agonist or antagonist of the receptor and whether the compound induces degradation of the receptor.

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An Optimized ChIP-Seq Protocol to Determine Chromatin Binding of Estrogen Receptor Beta,, however, have been more difficult, in part due to a lack of endogenous expression in cell lines and lack of specific antibodies. In this chapter, we provide an optimized stepwise ChIP protocol for a well-validated ERβ antibody, which is applicable for ChIP-Seq analysis of cell lines with exogenous expression of ERβ.

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Expression Profiles of Estrogen-Regulated MicroRNAs in Cancer Cells,or 1. This chapter details how to prepare cells for optimal estrogen response, directions for estrogen treatment, RNA extraction, different microRNA profiling approaches, and subsequent confirmations.

Carcinogenesis

,Immunoblot Detection of the Phosphorylation of the Estrogen Receptor α as an Outcome of GPR30 /GPERd subsequent immunoblotting techniques. In particular, phosphorylation of the ERα is one possible outcome of activation of the putative membrane estrogen receptor (mER), GPR30 or GPER1. Hence, phosphorylation represents a crosstalk event between GPR30 and ERα and may be important in estrogen-regulated physiology.

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Extraction of RNA and Analysis of Estrogen-Responsive Genes by RT-qPCR,bust and allows the measurement of gene expression in a variety of tissues and cells. This chapter describes the protocols used for RNA extraction and analysis as well as for RT-qPCR assay using hydrolysis (TaqMan-type) probes.

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1064-3745 expertsThis fully updated collection includes chapters on a wide array of techniques that are vital for advancing our understanding of the physiological and pathological effects of estrogen, mediated by estrogen receptor α (ERα) and estrogen receptor β (ERβ) as well as the non-genomic membrane-boun

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https://doi.org/10.1007/978-3-031-44294-0d subsequent immunoblotting techniques. In particular, phosphorylation of the ERα is one possible outcome of activation of the putative membrane estrogen receptor (mER), GPR30 or GPER1. Hence, phosphorylation represents a crosstalk event between GPR30 and ERα and may be important in estrogen-regulated physiology.

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The Italian Communist Party and Culturebust and allows the measurement of gene expression in a variety of tissues and cells. This chapter describes the protocols used for RNA extraction and analysis as well as for RT-qPCR assay using hydrolysis (TaqMan-type) probes.