Figure 3

Figure 3

AP2M1 increased drug resistance. A) Putative drug resistance distribution for Idarubicin, Daunorubicin, and Cytarabine in malignant HSPCs of scRNA‐seq data. The upper graph displays the distribution of drug resistance scores, and the lower compares the extent of drug resistance according to AP2M1 expression. The significance of the difference between groups was compared using the Wilcoxon rank‐sum test. The significance is as follows, ^*p < .05, ^**p < 0.01, ^***p < .001. B,C) Analysis of cell viability (B) and proliferation (C) in AP2M1‐overexpressing cells in response to idarubicin treatment. Control and AP2M1‐overexpressing cells were treated with idarubicin, and cell viability and proliferation were assessed using Cyto X staining. D) Fold change in the sub‐G1 fraction following idarubicin treatment in control and AP2M1‐overexpressing cells. Control and AP2M1‐overexpressing cells were treated with 10 µM idarubicin for 24 h, followed by PI staining. Cell cycle distribution was analyzed by flow cytometry. E,F) Assays evaluating changes in apoptosis‐related genes, including BCL2, BAX, caspase‐3 (CASP3), and caspase‐9 (CASP9). These alterations were assessed in both control and AP2M1‐overexpressing cells, as well as following drug treatment, at mRNA (E) and protein levels (F). G) Expression level of genes negatively regulating apoptotic signaling pathway (GO, 2001234). The scores were calculated using transcriptional profiles of HSPC from scRNA‐seq data. H) FACS assay with FITC‐annexin V/PI staining. The x‐axis represents the fluorescence intensity of FITC‐annexin V, indicating the binding of Annexin V to phosphatidylserine on the outer leaflet of the cell membrane, whereas the y‐axis shows the fluorescence intensity of PI, which penetrates cells with compromised membranes and stains their DNA. I) DiI‐stained images in the control and CMV‐ap2m1a group. Tumors in the DiI‐stained control group and CMV‐ap2m1a group were observed using fluorescence microscopy. Solid arrows indicate the presence of tumors, while hollow arrows denote the disappearance of tumors. J) qPCR analysis of CD41 mRNA expression level in zebrafish embryos injected with AML1‐ETO mRNA at the one‐cell stage and analyzed at 72 hpf. K) qPCR analysis of CD41 mRNA expression level in zebrafish embryos injected at the one‐cell stage with AML1‐ETO mRNA alone or co‐injected with AML1‐ETO and AP2M1 mRNAs. Embryos were treated with idarubicin (10 µM) from 24 to 72 hpf, and CD41 expression was compared between idarubicin‐treated and untreated groups within each injection condition.