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. 2015 Oct;20(9):1132–1141. doi: 10.1177/1087057115594187

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© 2015 Society for Laboratory Automation and Screening

This article is distributed under the terms of the Creative Commons Attribution-NonCommercial 3.0 License (http://www.creativecommons.org/licenses/by-nc/3.0/) which permits non-commercial use, reproduction and distribution of the work without further permission provided the original work is attributed as specified on the SAGE and Open Access page (http://www.uk.sagepub.com/aboutus/openaccess.htm).

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Figure 4. — Screening of lncRNA-protein interactions. (A) The tolerance of the assay to DMSO was tested over a 100-fold range (0.1%−10% final DMSO concentration) for both lncRNAs of interest. The lncRNAs, EZH2, and AlphaScreen assay components were found to be stable in DMSO up to 1%, well below the 0.1% DMSO concentration used for screening. Data represent triplicates of a single experiment that was repeated 3 independent times. Scatterplot of Prestwick Phytochemical library (Prestwick Chemical, Illkirch-Graffenstaden, France) screen for (B) BDNF-AS EZH2 (Z-factor = 0.65, 3.1% hit rate) and (C) HOTAIR-EZH2 (Z-factor = 0.85, 1.25% hit rate). A total of 320 compounds (10 µM, 0.1% DMSO) were tested. Data are the average of duplicates expressed as percentage inhibition relative to a positive control (no lncRNA in the reaction, set to 100% inhibition). The negative control (0% signal inhibition) consisted of lncRNA and protein in 0.1% DMSO. Red circles in (B) and (C) indicate biotin, which shows greater than 100% inhibition in the screen, confirming our data and serving as a positive control for future studies.