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Figure 5
(A) CSNK2B KD in CMS decreases BFU-E, and CSNK2B OE in non-CMS increases BFU-E, suggesting its critical role in regulating erythropoiesis. **P < 0.001. (B) Effect of CK2 inhibitor on CMS cells. TBB decreases BFU-E colonies in CMS cells in a dose-dependent manner. *P < 0.05; **P < 0.001. (C) Effect of CK2 inhibitor on CMS cells. CX4945 decreases BFU-E colonies more drastically in the CMS cells in a dose-response manner. **P < 0.01; ***P < 0.001; ****P < 0.0001. (D) CSNK2B KD results in major expression changes of critical TFs. qPCR results confirm decreased expression of TAL1, KLF1, RUNX1, IKAROS, and GATA1. *P < 0.05; ***P < 0.001. (E) GATA1 expression levels were altered significantly by modulation of CSNK2B levels in CMS and non-CMS cells under hypoxia. Graph shows GATA1 expression as measured by qPCR in (a) CMS cells, (b) CMS cells with CSNK2B KD, (c) CMS cells treated with CK2 inhibitor, (d) non-CMS cells, and (e) non-CMS cells with CSNK2B-OE. *P < 0.05; **P < 0.01. (F) CSNK2B regulates erythropoietic response through GATA1. Graph shows the effect of CSNK2B and GATA1 modulation on colony-forming potential of CMS and non-CMS cells. GATA1 OE partially rescues the erythropoietic suppression caused by CSNK2B in CMS. Further, KD of GATA1 in non-CMS results in loss of excessive erythropoiesis caused by OE of CSNK2B. *P < 0.05; **P < 0.001. For all the experiments (colony-forming assays as well as qPCR), iPSC-derived CD34+ cells were used. n = 3 per group. For A–F, 1-way ANOVA was performed, followed by multiple comparisons by Tukey’s test. For D, t tests were performed for each comparison.