Fig. 2: Erythroid-biased differentiation and increased self-renewal capacity of high-risk MDS-iPSC containing a C/EBPα bZIP frameshift mutation.

A Bar graph showing the total number of HPCs, percentage of early hematopoietic population (CD43⁺) and late hematopoietic population (CD34⁺ CD45⁺) on day 14 of differentiation. Total number of HPCs for normal and C22 taken from Fig. 1C. Statistical results are presented as mean ± SEM. ns = no significant, One-way ANOVA with Dunnett’s multiple comparisons. N = 4 independent experiments. Source data are provided as a Source Data file. B Percentage of hCD45⁺ cells (Left graph) engrafted in humanized niches implanted in NSG-SGM3 mice at 12 weeks. Each point represents an individual scaffold; each sample was transplanted into two mice, with technical replicates shown. Box plots display the full range of values (minimum to maximum), with the median indicated by a horizontal line. Lineage distribution (Right graph) within hCD45⁺ cells recovered from humanized niches in NSG-SGM3 mice. Bars represent mean values, and error bars indicate the standard error of the mean (SEM). Source data are provided as a Source Data file. C Total number of CFUs from 10⁴ –iPSC-HPCsgrown for 14 days in semi-solid medium. Statistical results are presented as mean ± SEM. ****p < 0, ***p < 0.0001, **p < 0.001, *p < 0.05 and (ns, no significant). One-way ANOVA with Dunnett’s multiple comparisons. N = 4 independent experiments. Exact p-values can be found in the Supplementary Data Table 5. Source data are provided as a Source Data file. D Number of each type of CFUs after 14 days in semisolid medium. Mean and SEM of different lines are shown. N = 4 independent experiments. Source data are provided as a Source Data file. E Relative percentage of each type of CFUs for 1 × 10⁴ of HPCs after 14 days in semisolid media. Statistical results are presented as mean ± SEM and ****p < 0.0001 and *p < 0.05, Two-way ANOVA with Dunnett’s multiple comparisons. N = 4 independent experiments. Exact p-values can be found in the Supplementary Data Table 5. Source data are provided as a Source Data file. F Representative flow cytometry panels showing the percentage of erythroblasts cells (blue square) (CD71+ CD235a+), immature myeloid cells, orange square (CD33⁺ CD11b⁺), and mature myeloid cells, green rectangle (CD11b+) obtained from colony assays. N = 3 independent experiments. Source data are provided as a Source Data file. G Fraction of erythroblasts (CD71⁺ CD235a⁺), immature myeloid (CD33⁺ CD11b⁺) and mature myeloid cells (CD11b+). Mean and SEM are shown. **** p < 0.0001, One-way ANOVA with Dunnett’s multiple comparisons. N = 3 independent experiments. Source data are provided as a Source Data file. H Diff-quick stained cytospins from colony assay showing erythroid cells (Red arrows), granulocytes (Green arrows) and monocytes (Pink arrows). The pictures were taken with a Leica DM6000 at ×40, 20 μm scale bar. N = 3 independent experiments. I Number of CFUs obtained from control, WT clones and mutant clones after second and third re-plating, each maintained for 14 days. Mean and SEM are shown. ****p < 0.0001, *p < 0.05, Two-way ANOVA with Dunnett’s multiple comparisons. N = 4 independent experiments. Exact p-values can be found in the Supplementary Data Table 5. Source data are provided as a Source Data file.