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. 2021 May 20;12(6):515. doi: 10.1038/s41419-021-03801-w

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© The Author(s) 2021

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. 3 — A, D Micrographs of the results of the primary and secondary tumor formation assays in the AA (A) and AW (D) sarcoma lines with EMX1 or EMX2 overexpression or in the BG cell line with EMXs downregulated by shRNAs (G). B, E, H Average number of primary (black bars) and secondary (gray bars) tumorspheres in AA (B), AW (E), and BG (H). In C, F, I, the mean sizes of the primary (bars in black) and secondary (bars in gray) tumors of AA (C), AW (F), and BG (I) are shown. J, M, P Representative images of tumor formation from isolated cells in the AA (J), AW (M), and BG (P) cell lines with different levels of EMX proteins. Sphere-formation efficiency of single cells for AA (K), AW (N), or BG (Q). Size of tumorspheres formed from an isolated cell in AA (L), AW (O), or BG with different levels of EMX proteins. All bars correspond to 500 µm. The mean of three independent experiments is represented in triplicate ± SD. Statistical analysis was performed with Student’s t-test (*p < 0.05; **p < 0.01; ***p < 0.001). S, T, V Effect of EMX1 and EMX2 levels on the percentage of CD133+ cells in sarcoma cell lines with different levels of EMX proteins. Graphs of the percentage of expression of the CD133 marker (conjugated with PE: CD133-PE), measured by FACS in the different models of lines AA (S) and AW (T) or BG (V) with different levels of EMX proteins.