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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 induces cell death with features of apoptosis and loss of membrane potential in MPNST cell lines. (a–d) ST88-14 (a,c) and T265-2c cells (b,d) were treated with WP1130 at the concentration of 2.5 µM, or transfected either with non-targeting (NT)-siRNA or Usp9X-siRNA. (a,b) Staining for annexin V/Propidium Iodide was performed prior to flow cytometric analysis. Representative flow plots are shown. N = 3. (c,d) Cells were stained with Mitochondrial Membrane Potential Assay Kit (TMRE), then flow cytometric analysis was performed. N = 3. (e,f) ST88-14 cells were treated with WP1130 at the concentration of 2.5 µM (e) or transfected either with non-targeting (n.t.)-siRNA or Usp9X-siRNA (f), then caspase 3-like enzymatic activity was measured at different time points. Column: mean. Error bar: SD, n = 3. * = 0.0209 ** = 0.0026 *** ≤ 0.001. (g) T265-2c cells were treated with WP1130 at the concentration of 2.5 µM prior to caspase 3-like activity measurement at different time points.