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. Author manuscript; available in PMC: 2025 Jul 1.
Published in final edited form as: Br J Haematol. 2024 Jun 12;205(1):207–219. doi: 10.1111/bjh.19563

Figure 6.

Figure 6.

(A-B) The Venn graph depicts the number of differentially expressed genes between cells expressing sWT1+/− (A) or sWT1−/− (B) and cells expressing other sWT1 transcripts and an empty control vector. (B) RNAseq heatmap indicating differentially expressed genes between sWT1+/− and other transcripts. The average expression of the indicated genes in primary leukemia samples from the BeatAML cohort is also displayed. (C) The competitive growth assay plot demonstrates a growth disadvantage of HOXA3 expressing cells (Dsred+) determined by flow cytometry. (D) Representative flow cytometry plots showing decreased CD71 expression in OCIAML3 and MOLM13 cells transduced expressing HOXA3 (Dsred+). (E) The competitive drug graphs depict increased resistance of HOXA3 expressing cells indicated by increased percentages of Dsred+ cells treated with dose gradients of cytarabine in the presence of doxycycline for 48 hours measured by flow cytometry. (F) The competitive growth assay plot shows a growth disadvantage of cells expressing GATA2. (G) Representative flow cytometry plot showing decreased CD71 expression in OCIAML3 and MOLM13 cells transduced with a GATA2 overexpression vector. (H) The competitive drug graphs depict the percentages of GATA2 expression cells (MOLM13, OCIAML3) treated with dose gradients of cytarabine and venetoclax for 48 hours measured by flow cytometry. (I) The representative immunoblot images showing increased p21 (encoded by CDKN1A) in Dox inducible sWT1−/− overexpressing cells in the presence of Dox for 3 days. (J) RT-qPCR data showing 2^-ΔΔCt changes of HOXA3, GATA2, and WT1 in sWT1+/−, HOXA3, GATA2, or HOXA3 overexpressing cells normalized to HPRT and empty vector control cells. (K) Representative cell cycle histograms depict percentages of cells in different cell cycle stages in HOXA3 or GATA2 overexpressing (Dsred+) or control (Dsred−) cells. The experiments were performed from 2-3 cell lines (OCIAML3, MOLM13, and MOLM14). (L) The graph demonstrates low expression of HOXA3 in CMK and K562 cells compared to other leukemia cell lines. Relative mRNA expression was normalized to HPRT control. (M) The competitive drug curves depict percentage changes of HOXA3, sWT1+/− and GATA2 overexpressing (DsRed+) K562 and CMK cells treated with a gradient dose of cytarabine.