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. 2021 Aug 31;36(2):438–451. doi: 10.1038/s41375-021-01394-z

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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. 6 — A Immunoblot analysis of USP15, KEAP1, NRF2, and ACTIN/GAPDH in MV4;11 and OCI-AML3 cells treated with vehicle (DMSO, 0 μM) or the indicated concentrations of USP15 inhibitor for 24 h. Data reflects three independent assays. B Left; histogram flow plots of DCFDA analysis of cellular ROS in the indicated AML cells treated for 24 h with DMSO (gray) or 10 μM of USP15 inhibitor (green) with the raw MFI values set shown. Right; the normalized mean-fluorescent intensity of DCFDA in the AML cell lines versus their DMSO controls. A parametric, ratio-paired t test was performed. *P = 0.01. The centerline depicts the median. Data is from four independent cell lines. C Relative NQO1 expression in the indicated AML cell lines treated with vehicle (DMSO) or USP15 inhibitor (10 μM) for 24 h (n = 3 replicates). D Cell-Titer Glo relative luminescence was measured at 72 h with USP15 inhibitor in the indicated AML cell lines. Luminescence detected in inhibitor-treated cells was normalized to the vehicle control cells treated with DMSO (n = 3 replicates per dose). The IC50 values for each cell line are shown. E Total number of colonies formed by MV4;11, OCI-AML3, and MOLM14 cell lines in methylcellulose after 7 days of treatment with the USP15 inhibitor (10 μM). (n = 3 per group). Data reflects two independent assays. F The total number of colonies formed by three independent patient-derived-xenografted (PDX) AML samples and normal CD34+ cells after 7–10 days of treatment with the USP15 inhibitor (10 μM). *P < 0.05. G Representative colony images from healthy CD34+, OCI-AML3 leukemic cell line, and three PDX AML samples treated with DMSO or USP15 inhibitor.