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. 2018 Jul 10;9:2668. doi: 10.1038/s41467-018-05127-2

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© The Author(s) 2018

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 license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license 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 license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 2 — Compound validation via in vitro assays. a Inhibition of SOD1^G93A-Derlin-1 interaction by #56 analogs in the in vitro immunoprecipitation (IP) assay. Purified SOD1^G93A-Derlin-1 complex form HEK293A cell lysates transfected with Flag-SOD1^G93A and Derlin-1-HA were incubated with the indicated compounds for 16 h. Then, the complex was immunoprecipitated using anti-Flag beads, followed by immunoblotting (IB) analysis with the indicated antibodies. b IC₅₀ values of notable #56 analogs and #56 in the in vitro assay. IC₅₀ values were calculated by generating standard curves. c, d Chemical structure of #56-20 and #56-26