Fig. 2

Yap Signaling Impacts HSPC Production in the Zebrafish Embryo (A) Whole-mount in situ hybridization (WISH) for runx1/cmyb in control and yap1mw48 mutants at 36 hpf. Scale bar, 100 μm. (B) Qualitative phenotypic distribution plot of embryos in (A) scored with low, medium, or high runx1/cmyb expression in the VDA at 36 hpf (n = 32 control, 33 yap1mw48; chi-squared test, ∗∗∗p < 0.001). (C) Whole-embryo qPCR on control and yap1mw48 mutants at 36 hpf (unpaired Student’s t test, ∗p < 0.05, ∗∗∗p < 0.001. Error bars indicate SEM). (D) WISH for runx1/cmyb at 36 hpf following heat-shock-mediated overexpression of constitutively active Yap (YapS87A, hs:Yap) at 12 hpf. Scale bar, 100 μm. (E) Distribution plot of embryos in 2D (n = 53 control, 48 hs:Yap; chi-squared test, ∗∗∗p < 0.001). (F) qPCR in 36 hpf embryos following Yap-OE at 12 hpf (unpaired Student’s t test, ∗p < 0.05, ∗∗∗p < 0.001. Error bars indicate SEM). (G) In vivo imaging of CD41:GFP in the CHT at 72 hpf after YAP-OE. Scale bar, 200 μm. (H) Quantification of fluorescent images in (G) (n = 9 control, 7 hs:Yap; unpaired Student’s t test, ∗p < 0.05. Error bars indicate SD). (I) rag1 expression in the thymus at 120 hpf with Yap-OE (n = 21). Scale bar, 100 μm. (J) Quantification of thymic area in (I) (n = 8; unpaired Student’s t test, ∗p < 0.05. Error bars indicate SD). (K) WISH for runx1/cmyb at 36 hpf following Yap-OE and sih morpholino (MO). The black bracket denotes VDA and arrowhead indicates uncleared primitive erythrocytes in the vein due to lack of blood flow. Scale bar, 100 μm. (L) Distribution plot of embryos in (K) (n ≥ 30 per condition; chi-squared test, ∗∗∗p < 0.001). (M) Quantification of runx1/cmyb staining intensity in (K) (n ≥ 30 per condition; two-way ANOVA and Tukey’s multiple comparison test, ∗p < 0.05, ∗∗∗p < 0.001. Error bars indicate SD).