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. 2006 Dec 7;35(2):e10. doi: 10.1093/nar/gkl1045

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© 2006 The Author(s).

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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In vitro transcription assay using plasmids pE3SP-shLuc and pU6-shLuc in nuclear extracts. Assays were done in absence or presence of α-amanitin, a specific inhibitor of Pol II- and Pol III-dependent transcription. (A) The expected band of 60 nt, corresponding to the expressed shRNA. (B) Additional fragment of 150 nt in length, resulting from pU6-shLuc (Pol III-dependent) transcription. (C) 5S rRNA transcript (Pol III-dependent), used as a positive control for assay, and inhibition by α-amanitin. Asterisk indicates an unspecific signal, inherent to the transcription from nuclear extract of FM3A cells.