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. 2018 Jul 2;8:9966. doi: 10.1038/s41598-018-27767-6

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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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Imatinib sensitivity in parental cells and T315I-acquired sublines of three cell lines. (a) Phosphorylation of CRKL in parental cells (upper panel) and T315I-acquired subline (#1) (lower panel) of K562 (left), TCCS (middle), and KOPM28 (right). Cells were cultured in the absence or presence of 1 μM for 24 hours. Blue shade, red line, and blue line indicate isotype control, phospho-CRKL of untreated cells, and phospho-CRKL of imatinib-treated cells, respectively. (b) Flow cytometric analysis of PI staining in parental cells (upper) and T315I-acquired subline (#1) (lower panel) of K562 (left), TCCS (middle), and KOPM28 (right). PI staining was performed after 24 hours culture in the absence (upper) or presence (lower) of 1 μM of imatinib. Percentages of cells in sub-G0/G1, G0/G1, S, and G2/M phases are indicated in each panel. (c) Flow cytometric analysis of apoptotic cell death in parental cells (upper) and T315I-acquired subline (#1) (lower panel) of K562 (left), TCCS (middle), and KOPM28 (right). Cells were cultured in the absence or presence of 1 μM for 24 (TCCS and KOPM28) or 60 (K562) hours, and analyzed with Annexin V-binding (horizontal axis) and 7AAD-staining (vertical axis) using flow cytometry. The percentages of living cells and early and late apoptotic cells are indicated. (d) Dose-response curve of imatinib sensitivity in parental cells (blue line) and T315I-acquired sublines (#1 and #2) (red and green lines, respectively) of K562 (upper left), TCCS (upper right), and KOPM28 (lower left). The vertical axis indicates % viability in the alamarBlue cell viability assay and the horizontal axis indicates log concentration of imatinib (nM).