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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. 6 — a, Western blot assay to quantify RBFOX2 knockdown efficiency in NB4 cells. b-c, Flow cytometric analysis of CD11b⁺ cell populations (b), and relative gene expression of CD14 and CD11b with qPCR quantification (c) in control (shNS) and RBFOX2 KD (shRBFOX2-1 and shRBFOX2-2) NB4 cells after treatment with 500 nM ATRA for 72 hr as compared to DMSO-treated cells. n = 3. d, Western blot assay to quantify RBFOX2 knockdown efficiency in MOLM13 cells. e-f, Flow cytometric analysis of CD11b⁺ cell populations (e) and CD11b mRNA expression level (f) in control (shNS) and RBFOX2 KD (shRBFOX2-1 and shRBFOX2-2) MOLM13 cells. n = 3. g, Wright-Giemsa staining of cytospin slides of control (shNS) and RBFOX2 KD (shRBFOX2-1 and shRBFOX2-2) MOLM13 cells. Arrows indicate differentiated cells. Scale bar, 20 µm. h, Western blots showing the knockdown efficiency of RBFOX2 in human cord blood-derived CD34⁺ (CB-CD34⁺) cells. i, Flow cytometric analysis of the percentage of CD34⁺ cell population in control (shNS) and RBFOX2 KD (shRBFOX2-1 and shRBFOX2-2) CB-CD34⁺ cells. Day 2 and Day 5 represent the number of days post-transduction. j, Quantification of the percentage of different hematopoietic lineage progenitor colonies in control (shNS) and RBFOX2 KD (shRBFOX2-1 and shRBFOX2-2) CB-CD34⁺ cells. n = 4. P values were calculated by two-way ANOVA, Dunnett’s multiple comparisons test. k-n, Flow cytometric analysis of megakaryocyte (k), erythroid (l), granulocyte (m) and monocyte (n) differentiation of CB-CD34⁺ cells (five days post-transduction) transfected with shNS or RBFOX2 shRNAs (shRBFOX2-1 and shRBFOX2-2). n, biologically independent samples. Data are presented as mean values +/− SEM. For c and f, two-sided P values by Student’s t-test.

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