Fig. 5: Erythroid differentiation of high-risk MDS-iPSC containing a C/EBPα bZIP frameshift mutation reveals erythrodysplasia even after 5-Aza treatment. | Nature Communications

Fig. 5: Erythroid differentiation of high-risk MDS-iPSC containing a C/EBPα bZIP frameshift mutation reveals erythrodysplasia even after 5-Aza treatment.

From: A heterozygous CEBPA mutation disrupting the bZIP domain in a RUNX1 and SRSF2 mutational background causes MDS disease progression

Fig. 5

A Percentage of early erythroid progenitor cells (CD71+), erythroblasts (CD71+ CD235a+) and mature erythrocytes (CD235a+) during several time points of erythroid differentiation. Mean and SEM are shown. **** <0.0001, **p < 0.001, *p < 0.05 and (ns, no significant), Two-way ANOVA with Dunnett’s correction. 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. B Bar graph showing CD71 geometric mean (gMFI) of each cell line at day 18 of erythroid differentiation. Mean and SEM are shown. * p < 0.05, **p < 0.001. 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. C Diff-quick stained cytospins showing erythroid dysplasia; Giant erythroblast (black arrow), Mitotic erythroblast (Grey arrows) in MDS27-C22.7 (High-risk-1) and MDS27-C22.20 (High-risk-2), Multinucleated erythroblast (green arrows), bi-nucleated erythroblast (blue arrows) and nuclear bridge (pink arrows). The pictures were taken with a Leica DM6000 microscope at ×100 magnification, 20 μm scale bar. N = 4 independent experiments. D Bar graph represents the percentage of the aberrant cells of MDS27 clones relative to the aberrant morphology of hiPSC control. Results are presented as mean ± SEM and ****p > 0.0001, Two-way ANOVA with Dunnett’s multiple comparisons. N = 4 independent experiments. Source data are provided as a Source Data file. E Bar graph represents the number of CFUs obtained from control, MDS27-C22 (Low-risk), and MDS27-C22.7 (high-risk) in the presence of different concentrations of 5-Aza or vehicle control (DMSO). Mean and SEM are shown. ****p < 0.0001, **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. F Diff-quick stained cytospins from colony assay showing aberrant morphology in MDS27- C22.7 (high risk) after 5-Aza treatment; nuclear budding early orthochromatic erythroblasts (red arrow), bi-nucleated pronormoblast (green arrow), multinucleated basophilic erythroblast (blue arrows) and binucleated orthochromatic (nuclear bridge) (pink arrows). The pictures were taken with a Leica DM6000 microscope at ×63 magnification, 40 μm scale bar. N = 3 independent experiments. Source data are provided as a Source Data file. G May–Grunwald Giemsa stained bone marrow smears from MDS27 patient after receiving 3 rounds of Aza treatment showing aberrant erythroid cells. The pictures were taken with a Leica DM6000 at ×63 magnification. H Number of CFUs obtained from CD34+ sorted cells from MDS27 patient samples in normoxia (before and after disease progression) and hypoxia (after disease progression). Cell were maintained for each replating. Source data are provided as a Source Data file.

Back to article page