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Fig. 5 Ddx41 regulates HSPC number via the cGAS-STING inflammatory pathway (A) Schematic of the cGAS-STING pathway knockdown experiments with HSPC quantification. (B) Flow cytometry plots of cd41:gfp+ HSPCs from ddx41 mutants injected with control morpholino (left), splice-blocking sting morpholino (middle), or translation-blocking (ATG) cgas morpholino (right) at 40 hpf. (C) Graph depicting the absolute number of cd41:gfp+ cells per embryo shown in (B). (D) Schematic of the STING knockdown and RNASEH1 overexpression experiment with HSPC quantification. (E) Flow cytometry plots of runx1:mcherry+ HSPCs from Tg(hsp70:M27RNASEH1-GFP)-negative (top left) and Tg(hsp70:M27RNASEH1-GFP)-positive (top right) uninjected ddx41 mutants versus Tg(hsp70:M27RNASEH1-GFP)-negative (bottom left) and Tg(hsp70:M27RNASEH1-GFP)-positive (bottom right) STING morpholino-injected ddx41 mutants at 40 hpf. (F) Graph depicting the number of runx1:mcherry+ HSPCs per embryo from (E). Graphs display means ± stds with p values calculated with a one-way ANOVA with Tukey’s multiple testing correction, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001, ns, not significant (p > 0.05). N = 3–6 replicates per experiment.
Reprinted from Developmental Cell, 56(5), Weinreb, J.T., Ghazale, N., Pradhan, K., Gupta, V., Potts, K.S., Tricomi, B., Daniels, N.J., Padgett, R.A., De Oliveira, S., Verma, A., Bowman, T.V., Excessive R-loops trigger an inflammatory cascade leading to increased HSPC production, 627-640.e5, Copyright (2021) with permission from Elsevier. Full text @ Dev. Cell