Fig. 4

Fig. 4 YAP Activity Is Required for Maintenance, but Not Acquisition, of Hematopoietic Commitment in HE (A) WISH for runx1, cmyb, and gata2b at 24 hpf in WT and yap1mw48 mutants. Black arrowheads indicate putative HE cells expressing cmyb in VDA versus primitive erythrocytes in the vein not yet mobilized by circulation. Scale bar, 100 μm. (B) Distribution plots of embryos in (A) (n ≥ 30 per condition). (C) WISH for runx1, cmyb, and gata2b at 30 hpf in WT and yap1mw48 mutants. (D) Distribution plots of embryos in (C) (n ≥ 30 per condition; chi-squared test, ∗∗∗p < 0.001). (E) WISH for runx1/cmyb at 36 hpf upon Yap-OE at 12 hpf and treatment with DMSO or DAPT from 12–36 hpf. Scale bar, 100 μm. (F) Distribution plot of embryos in (E) (n ≥ 30 per condition; chi-squared test, ∗∗∗p < 0.001). (G) Quantification of runx1/cmyb staining intensity in (F) (n ≥ 30 per condition; two-way ANOVA and Tukey’s multiple comparison test, ∗∗p < 0.01, ∗∗∗p < 0.001. Error bars indicate SD). (H) Max proj. of confocal z stack of Flk1:GFP embryos (cyan) at 36 hpf following Yap-OE. EDU+ nuclei shown in magenta. Green arrowheads denote dorsal EDU+ endothelial nuclei in the aorta. Yellow arrowheads indicate Flk1+ EDU+ cells in the VDA and perivascular space between the DA and PCV. Scale bar, 50 μm. (I) Quantification of Flk1+ EDU+ cells in (H) (n = 20; unpaired Student’s t test, ∗∗∗p < 0.001. Error bars indicate SD).