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. 2013 May 9;2(1):214. doi: 10.1186/2193-1801-2-214

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© Villablanca et al.; licensee Springer. 2013

This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Dihydrotestosterone (DHT) treatment down regulates P450 aromatase expression and protein levels, and reduces estradiol production from murine ERα +/+ and ERα −/− male aortic endothelial cells. Aortic endothelial cells isolated from male ERα +/+ and ERα −/− mice were treated with DHT (1 nM or 1 uM) for 48 hours. Following treatment, total RNA was isolated and used for qRT-PCR analysis of expression of the P450 aromatase gene. Gene expression levels were normalized to GAPDH and expressed as percent change compared to control (panels A and B). To ascertain the amplification and integrity of the resulting qRT-PCR products, the products were separated on a 2% agarose gel (inset, top panel: P450 aromatase and bottom panel: GAPDH). Cell supernatants were collected following treatment and used for ELISA analysis of estradiol release (panels C and D). Results are expressed as means ± SEM. ** p < 0.01 and * p < 0.05 compared to vehicle control.