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. 2023 Aug 28;25(9):1359–1368. doi: 10.1038/s41556-023-01213-w

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© The Author(s) 2023

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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Extended Data Fig. 7 — a, Western blot of RBFOX2 protein in normal controls (human bone marrow CD43⁺) and AML cell lines. b, Kaplan-Meier survival analysis in TCGA-AML dataset (n = 106) using GEPIA. The patients were divided into two groups of equal size based on RBFOX2 levels. n, biologically independent patients. P value was detected by the log-rank test. c-d, Relative Rbfox2 gene expression quantified by qPCR (c, n = 3) and western blot assay (d) in control (shNS) and Rbfox2 KD (shRbfox2-1 and shRbfox2-2) mouse MLL-AF9 (MF9) cells. e-f, Image (e) and quantification (f, n = 2) of effects of Rbfox2 knockdown on the colony-forming of mouse MA9 AML cells. g, Flow cytometric analysis of LSC populations in mouse MA9 leukemia cells. h, Flow cytometric analysis of CD11b⁺ cell populations in mouse MA9 leukemia cells. i, Western blot of RBFOX2 protein in control (shNS) and RBFOX2 KD (shRBFOX2-1 and shRBFOX2-2) in in vivo bone marrow samples. j, Western blot showing RBFOX2 expression in control and RBFOX2 KD (shRBFOX2-1 and shRBFOX2-2) AML-PDX cells. k, Effects of RBFOX2 knockdown (shNS [n = 4], shRBFOX2-1 [n = 4] and shRBFOX2-2 [n = 3]) on AML-PDX cell growth. P value was calculated by one-way ANOVA, Dunnett’s multiple comparisons test. l, Flow cytometric analysis of CD11b⁺ cell populations in control and RBFOX2 KD AML-PDX cell (2017-129). m, Flow cytometric analysis of CD11b⁺ (top panel) or CD11b⁺ and CD15⁺ (bottom panel) cell populations in control and RBFOX2 KD AML-PDX cell (HBT22-0148). n, The hCD45 and hCD33 double positive cells (CD45⁺CD33⁺) were utilized to determine the engraftment of human AML-PDX cells (2017-129⁴⁴) in recipient mice on day 34 post xeno-transplantation (shNS [n = 8], shRBFOX2-1 [n = 7] and shRBFOX2-2 [n = 9]). o, Quantification of leukemia burden in control (shNS [n = 5]) and RBFOX2 KD (shRBFOX2-1 [n = 4] and shRBFOX2-2 [n = 3]) bone marrow (BM) cells. p, Relative gene expression of CD14 and CD15 in control (shNS [n = 4)) and RBFOX2 KD (shRBFOX2-1 [n = 4] and shRBFOX2-2 [n = 3]) BM cells. n, biologically independent mice. Data are presented as mean values +/− SEM. For c, n, o and p, two-sided P values by Student’s t-test.

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