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J.T. Weinreb et al.
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Figure 5. Ddx41 regulation of ATM/ATR signaling contributes to proper erythroid progenitor proliferation. (A) Cell cycle analysis of gata1:dsred+ erythroid cells from sibling controls (top) and ddx41 mutants (bottom) treated with dimethyl sulfoxide (DMSO) (left), 30 nM KU60019 (Ataxia-telengiectasia-mutated [ATMi] inhibitor, mid- dle), and 30 nM AZ20 (Ataxia-telengiectasia and Rad3-related [ATRi] inhibitor, right) after a 2-hour pulse of 5-ethynyl-2′-deoxyuridine (EdU) at 28 hours post fertiliza- tion (hpf). EdU incorporation (y-axis) and DAPI content (x-axis) were measured by flow cytometry at 30 hpf. (B) Quantification of the percentage of cells in each cell cycle phase from (A). (C and E) Flow cytometry plots of gata1:dsred+ erythroid cells from sibling controls (C) and ddx41 mutants (E) treated with DMSO (left), 30 nM KU60019 (ATM inhibitor, middle), and 30 nM AZ20 (ATR inhibitor, right). (D and F). Graphs depicting the absolute number of gata1:dsred+ erythroid cells per embryo from (C) and (E). Graphs display means ± standard deviations (stds) with P-values calculated with a one-way ANOVA with Tukey’s multiple testing correction, *P<0.05, ***P≤0.001, ****P≤0.0001. For flow cytometry, n=3-5 pools of ~5-20 embryos per pool.
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haematologica | 2022; 107(3)