Fig. 3: C/EBPα bZIP frameshift mutation on its own does not confer self-renewal capacity to healthy HPCs or to diseased HPCs after correction of SRSF2 and RUNX1 mutations.

A Bar graph showing the number of HPCs and percentage of early hematopoietic population (CD43+) and late hematopoietic population (CD34⁺ CD45⁺) on day 14 of differentiation. Statistical results are presented as mean ± SEM. ns = no significant, One way ANOVA with Dunnett’s multiple comparisons. N = 3 independent experiments. Source data are provided as a Source Data file. B Total number of CFUs from 10⁴ iPSC-HPC cells grown for 14 days in semi-solid medium. Statistical results are presented as mean ± SEM. *p < 0.05; **p < 0.01, t-test. One-way ANOVA with Dunnett’s multiple comparisons. N = 3 independent experiments. Exact p-values can be found in the Supplementary Data Table 5. Source data are provided as a Source Data file. C Number of each type of CFU after 14 days in semisolid medium. Mean and SEM of different lines are shown. N = 3 independent experiments. Source data are provided as a Source Data file. D Number of CFUs obtained from healthy iPSC, healthy iPSC CRISPR control, and healthy iPSC clones containing C/EBPa^bZIP-fs (C5 and C12) after first and second re-plating, each maintained for 14 days. Mean and SEM are shown. **p < 0.001, *p < 0.05 and (ns, no significant), Two-way ANOVA with Dunnett’s multiple comparisons. N = 3 independent experiments. Exact p-values can be found in the Supplementary Data Table 5. Source data are provided as a Source Data file. E Diff-quick stained cytospins from colony assay showing erythroid cells (Pink arrows), granulocytes (Red arrows), and monocytes (black arrows). The pictures were taken with a Leica DM6000 at ×40, 20 μm scale bar. N = 3 independent experiments. F Bar graphs showing the number of cells obtained after the first and second replating. Statistical results are presented as mean ± SEM. ns, no significant, t test. One-way ANOVA with Dunnett’s multiple comparisons. N = 3 independent experiments. Source data are provided as a Source Data file. G Bar graph showing the number of HPCs and percentage of early hematopoietic population (CD43⁺) and late hematopoietic population (CD34⁺ CD45⁺) on day 14 of differentiation derived from MDS27-C22.7 S/R reverted (C5 and C8). Statistical results are presented as mean ± SEM. ns = no significant, One-way ANOVA with Dunnett’s multiple comparisons. N = 4 independent experiments. Data from C22 and C22.7 is taken from Fig. 2A. Source data are provided as a Source Data file. H Total number of CFUs from 10⁴ iPSC-HPC cells grown for 14 days in semi-solid medium. Statistical results are presented as mean ± SEM. *p < 0.05; **p < 0.01, One-way ANOVA with Dunnett’s multiple comparisons. N = 3 independent experiments. Data from C22 and C22.7 is taken from Fig. 2B. Exact p-values can be found in the Supplementary Data Table 5. Source data are provided as a Source Data file. I Number of each type of CFUs after 14 days in semisolid medium. Mean and SEM of different lines are shown. N = 3 independent experiments. Data from C22 and C22.7 is taken from Fig. 2C. Source data are provided as a Source Data file. J Number of CFUs obtained from MDS27-C22.7 S/R reverted (C5 and C8) after first and second re-plating, each maintained for 14 days. Mean and SEM are shown. ****p < 0.0001, ***p < 0.001, **p < 0.01, Two-way ANOVA with Dunnett’s multiple comparisons. N = 3 independent experiments. Data from C22 and C22.7 is taken from Fig. 2H. Exact p-values can be found in the Supplementary Data Table 5. Source data are provided as a Source Data file. K Number of the single cells obtained after each replating. Statistical results are presented as mean ± SEM. ns= no significant ****p < 0.0001, ***p < 0.001, ns, no significant. One-way ANOVA with Dunnett’s multiple comparisons. N = 3 independent experiments. Data from C22 and C22.7 is taken from Supplementary Fig. 4C. Source data are provided as a Source Data file. L Fraction of erythroblasts (CD71+ CD235a+), immature myeloid (CD33+ CD11b+) and mature myeloid cells (CD11b+). Mean and SEM are shown. *p < 0.05; **p < 0.01; ****p < 0.0001; ns, no significant. One-way ANOVA with Dunnett’s multiple comparisons. N = 3 independent experiments. Exact p-values can be found in the Supplementary Data Table 5. Source data are provided as a Source Data file. M Diff-quick stained cytospins from colony assay showing erythroid cells (Pink arrows), and monocytes (Black arrows). The pictures were taken with a Leica DM6000 at ×63, 40 μm scale bar. N = 3 independent experiments.