Fig. 5: IGF2BP3 is highly expressed in LSCs/LICs and required for the maintenance of AML stemness.

A Expression of IGF2BP3 mRNA in AML patient samples and healthy donors from the TNMplot database. Box plot, center line, median; box limits, upper and lower quartiles; whiskers, 1.5× interquartile range. n = 407 healthy samples; n = 151 AML samples. B Expression of IGF2BP3 in CD34⁺ and CD34^- cells from BM of healthy donors (normal) or AML patients as detected by microarray (GSE30029). n = 31 CD34⁺ normal samples; n = 46 CD34⁺ AML samples; n = 44 CD34^- AML samples. C Expression of IGF2BP3 in blast (LSC⁻, non-engrafting) or LSC (LSC⁺, engrafting) cells from AML patients (GSE199452). n = 40 LSC⁻ samples; n = 70 LSC⁺ samples. D Experimental scheme for (E–G). Patient-derived CD34⁺ cells were transduced with shNS or shRNAs targeting IGF2BP3 and subjected to different assays. E–G Growth curves (E), percentages of Annexin V⁺ apoptotic cells (F), and colony numbers (G) of patient-derived CD34⁺ leukemia cells with or without IGF2BP3 KD. Bar = 200 μm. H The percentage of L-GMP population (CD45.2⁺Lin^-c-kit⁺Sca1^-CD34⁺CD16/32⁺) in BM of the primary BMT mice. n = 6 mice per group. I, J Statistics of the percentage of Annexin V⁺ (I) or Ki67⁺ (J) L-GMPs of mice in (H). n = 6 mice. K In vitro LDA assays using MA9-transduced HSPCs from Igf2bp3 WT and KO mice. Logarithmic plots show the percentage of nonresponding wells (those without any colonies) at different doses of cells seeded. The estimated LSC/LIC frequency is calculated by ELDA and shown. L, M In vitro LDA assays of mouse HSPCs co-transduced with MA9 plus shRNA targeting Igf2bp3 (L) or its target genes (M). N HSPCs collected from Igf2bp3 WT and HO mice were co-transduced with MA9 and target gene-overexpressing viruses and selected in methylcellulose medium with G418 and puromycin for 7 days before seeded for LDA assays. Mean ± SD values are shown. n = 2 biologically independent repeats in E, G while n = 3 biologically independent repeats in (F). Statistical analysis: Two-tailed student’s t-test (A, C, F, H–J); two-way ANOVA (B); One-sided Chi-squared test (K–N). Source data are provided as a Source Data file.