Figure 2.

D3 Negatively Regulates Hh-Mediated Vascular Niche Specification

(A) Gli:mCherry expression was decreased in D3-treated embryos compared to controls by WISH. Cyclopamine co-treatment caused synergistic reduction. Scale bar, 200 μm.

(B) Qualitative phenotypic distribution of embryos from (A) scored for gli:mcherry (n > 20 embryos/condition).

(C) Gli:mCherry FACS showed significant decreases in Hh responsive cells in D3 (54%, ^∗p = 0.006), cyclopamine (70%, ^∗∗p = 0.001), or co-treated embryos (91%,^∗∗∗p = 0.001) (5 embryos/sample × 4 replicates/condition). Error bars, mean ± SD.

(D) Qualitative phenotypic distribution of gli:mcherry expression in vdra morphants occurred in the presence or absence of D3 treatment (n > 20 embryos/condition).

(E) D3 reduced Notch reporter activity in intersomitic and axial vessels, similar to cyclopamine, by WISH. Co-treatment exacerbated Notch inhibition. Scale bar, 100 μm.

(F) Qualitative phenotypic distribution of embryos from (D) scored for Notch-GFP (n > 20 embryos/condition).

(G) SCL-GFP embryos treated with D3 (12–30 hpf) had reduced scl+ hemogenic endothelium by WISH for GFP. Scale as in (E).

(H) Qualitative phenotypic distribution of embryos from (F) scored for scl:GFP (n > 10 embryos/condition).

See also Figure S2.