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. 2022 Feb 15;12:2509. doi: 10.1038/s41598-022-06456-5

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© The Author(s) 2022

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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HTS results. (a) Raw FRET results for A455E NBD1 (acceptor and truncation at residues 487 and 654) screen using a 50,240 small molecule diversified library. Blue dots represent each averaged E_FRET value calculated based on two D and D + A pairs, and measured for compounds from 157 plates (1,536-well plate format, 600 beads/well, 320 compounds/plate). Black lines show ± 3SD for each plate. 133 primary hit compounds are indicated in orange circles (increase in FRET) or green circles (decrease in FRET) (0.26% hit ratio). Gray circles indicate averaged FRET values outside of ± 3SD but either or both single FRET values inside of ± 1.75SD. (b) Graph showing E_FRET values (mean ± SD, n = 32) of DMSO control for each plate. (c–e) Black dots represent coefficient of variation (SD/mean) of E_FRET values (c), or signal to background (mean D signal/mean background) (d) for each plate.