Endothelial Notch signaling acts upstream of Bmp to control PE formation. A and D, epi:GFP embryos at 60 hpf (A) or 80 hpf (D) either untreated or treated with LDN‐193189 or RO4929097 and immunostained for GFP (green), myosin heavy chain (MHC, red), pSmad1/5 (white) and nuclei counterstained with DAPI (blue). Top panels, 3D projections and lower panels optical sections. The DP was digitally isolated in 3D projections. Arrowhead, PE cluster. Arrows, epicardial cells. B, Quantification of PE cell number in A. C, Quantification of DP pSmad1/5+ cell number in A. E, Quantification of PE cell number in D. F, Quantification of DP pSmad1/5+ cell number in D. at, atrium; DP, dorsal pericardium; hpf, hours post fertilization; LDN, LDN‐193189; PE, proepicardium; RO, RO4929097; v, ventricle. DP digitally isolated in 3D projections. Scale bar: 50 μm. Data are means ± SD, unpaired two‐tailed Student's t‐test. ***P < .001, ns, nonsignificant
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