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Figure 3 The OFT endothelium is surrounded by smooth muscle cell progenitors expressing fibronectin and elastin. (A) Staining of Fibronectin1 (magenta) on Tg(myl7:GFP; fli1a:nls-mCherry), highlighting the myocardium (white) and the endothelium (green) at 72 hpf. Scale bar: 20 μm. V: ventricle. Fibronectin1 (anti-Fn1, magenta) counterstaining on Tg(kdrl:GFP) and Elastinb (anti-Elnb, magenta) counterstaining on Tg(fli:lifeact-EGFP) showing their expressions in the OFT at 72 hpf (B), (B’) respectively) at 96 hpf (C), (C’) respectively) and at 120 hpf (D, D’ respectively). Scale bar: 20 µm. Arrows show the Fn1 localisation within the valve leaflets. Results obtained from three independent experiments. (E) Scheme of the three layers shown in A’, B, C and D (magenta, smooth muscles; green, endothelium; grey, myocardium; Fibronectin1, magenta lines) at 72hpf, 96hpf and 120hpf.