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. 2010 Nov 1;137(21):3697–3706. doi: 10.1242/dev.051664

Fig. 6.

Fig. 6.

The midline-positive signals SHH and VEGF are necessary for dorsal aortae fusion independent of Chrd. (A) Whole-mount double in situ hybridization for Chrd (blue) and Cx40 (purple) in control embryo showing fused dorsal aortae at several somite levels anterior to Chrd downregulation in the notochord (n=10). (B) Transverse section of A showing fusing dorsal aortae under a Chrd(–) notochord. (C) As in A but treated with 100 μM cyclopamine (a hedgehog inhibitor) from stage 10 to stage 13, showing unfused dorsal aortae even though Chrd was downregulated in the notochord [as seen in control (n=9)]. (D) Transverse section of C. (E) As in C but treated with 10 μM Ki8751 (a VEGFR2 inhibitor) showing unfused dorsal aortae despite Chrd downregulation (n=8). (F) Section of E. (G,H) RT-PCR analysis of levels of Chrd, Vegf, Fgf2, Shh and Ptch1 in embryos treated with cyclopamine and Ki8751, respectively. (I) RT-PCR for Bmp2 and Bmp4 in cyclopamine- and Ki8751-treated embryos. Error bars indicate s.d. of two replicate experiments containing three pooled sibling embryos.