Fig. 4: High-risk MDS-iPSC containing a C/EBPα bZIP frameshift mutation exhibits a block in the granulocytic differentiation. | Nature Communications

Fig. 4: High-risk MDS-iPSC containing a C/EBPα bZIP frameshift mutation exhibits a block in the granulocytic differentiation.

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

Fig. 4

A Schematic representation of the myeloid differentiation protocol. Artwork generated with powerpoint Bundle-Biology. B Representative images of flow cytometric analysis of CD11b+ CD14− (granulocytes) and CD11b+ CD14+ (Monocytes) for the indicated cell types on day 0, day 4, and day 7 of differentiation. N = 4 independent experiments. Source data are provided as a Source Data file. C Percentages of CD11b+ CD14− (granulocytes) and CD11b+ CD14+ (Monocytes). Mean and SEM are shown ****p < 0.0001, *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 Diff-quick stained cytospins showing the presence of granulocytic cells (green arrows) and erythroblasts (red arrows). The pictures were taken with a Leica DM6000 microscope at ×40 magnification, 20 μm scale bar. N = 4 independent experiments. E Diff-quick stained cytospin showing aberrant morphology of myeloid cells (black arrows) in MDS27-C22 (Low-risk) and MDS27 C22 CRISPR control. The pictures were taken with a Leica DM6000 microscope at ×100 magnification, 20 μm scale bar. N = 4 independent experiments. F May–Grunwald Giemsa stained bone marrow smears from MDS27 patient at disease progression showing aberrant myeloid cells (from Supplementary Fig. 1A). The pictures were taken with a Leica DM6000 at ×63 magnification. G RNA expression of C/EBPα, RUNX1, SPI1 (PU.1), GATA2 and LMO2 measured by qRT-PCR, normalized against the GAPDH housekeeping gene and expressed relative to HPCs expression. N = 3 independent experiments. Mean and SEM are shown ****p < 0.0001, ***p < 0.001, **p < 0.01, *p < 0.05, and (ns, no significant), Two-way ANOVA with Tukey’s correction. 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.

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