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. 2019 Apr 5;9:5695. doi: 10.1038/s41598-019-42131-y

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© The Author(s) 2019

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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Decreased clonogenic and tumorigenic potential of mesenchymal DU 145 SKP2 KD cells is accompanied by a decrease of CD44⁺CD24⁻ cancer stem cell subpopulation. (A) Skp2 and p27^Kip1 expression in SKP2 KD cells were determined by western blot. Representative images are from three independent experiments. β-actin was used as a loading control. (B) Analysis of CD44 and CD24 markers in control and SKP2 KD cells using flow cytometry. Dot plots show representative results from three independent experiments. (C) Quantitation of results shown in (B). Data are presented as the mean ± SD of the percentage of CD44⁺CD24⁻ cancer stem-like cells (n = 3). (D) The capability of spheroid formation and growth in anchorage-independent conditions in SKP2 KD cells and control cells. (E) Quantitation of results shown in (D). Data are shown as the mean ± SD of the number of spheroids per well. Results are from three independent repetitions. (F) The xenograft formation rate of SKP2 KD cells in SHO mice. Tumour formation rate was then calculated for control and SKP2 KD cells. A number of mice per group, n = 10. (G) ALDH1 activity in DU 145 mesenchymal cells with Skp2 knocked down. Data are presented as the mean ± SD of the percentage of cells with active ALDH1. Results are from three independent experiments.