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. 2018 Nov 26;8:17390. doi: 10.1038/s41598-018-35806-5

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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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Usp9X inhibition causes massive reduction in cellular viability in MPNST cell lines. (a–c) ST88-14 (a), T265-2c (b), 90-8 cells (c) were treated with increasing concentrations of WP1130 for 72 h. Cellular viability was determined by CellTiter-Glo assay and the IC₅₀-values were calculated based on a non-linear regression analysis. Data are presented as mean and SD, n = 3. (d–f) Usp9X was silenced in ST88-14 (d), T265-2c (e), 90-8 cells (f) with non-targeting (NT) RNA or siUsp9X for 24, 48 and 72 h. Cellular viability was determined by CellTiter-Glo assay and relative cell viability was calculated. Data are presented as mean and SD, n = 3. * = 0.0129 *** ≤ 0.001. (g–j) Usp9X knock down was performed in ST88-14, T265-2c and 90-8 cells. Whole-cell extracts were examined by Western blot for Usp9X. ß-actin Western blot analysis was performed to confirm equal protein loading. Bar graphs show protein quantification analyzed through ImageJ. Data are presented as mean and SD, n = 3.