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Endocrine-disrupting chemicals (EDCs) such as 17-ethynylestradiol, 17-estradiol, estrone, and para-nonylphenol have been measured in wastewater-treatment plant effluents, surface waters, sediments, and sludge and have been shown to induce liver-specific vitellogenin (vtg) messenger RNA in male fathead minnows (Pimephales promelas). The purpose of the present study was to establish minimal concentrations of select EDCs necessary to induce transcription of vtg in 48-h static renewal exposures, as measured by quantitative real-time thermal cycle amplification. Adult males were exposed to 17-ethynylestradiol, 17-estradiol, estrone, and para-nonylphenol. Dose-dependent increases in vtg expression were significant with all chemicals tested. The lowest concentrations of these chemicals to induce measurable vtg expression, with significant difference from respective controls, were 17-ethynylestradiol, 2.2ngL(-1); para-nonylphenol, 13.9gL(-1); 17-estradiol, 42.7ngL(-1); and estrone, 46.7ngL(-1), measured as 48-h average concentrations. The present experiments were designed to frame a commonly acceptable approach for investigators who conduct static, in-laboratory proxy environmental aquatic exposures. The present study highlights the need for investigators to report in peer-reviewed submissions the observed concentration values for minimal induction levels when measuring molecular responses to chemical exposures by means of real-time polymerase chain reaction, quantitative polymerase chain reaction, or other omic technologies. Environ Toxicol Chem 2015;34:2796-2802. Published 2015 Wiley Periodicals, Inc. on behalf of SETAC. This article is a US Government work and is in the public domain in the United States of America.