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. 2000 Feb 1;105(3):261–270. doi: 10.1172/JCI8856

Figure 6.

Figure 6

Neuropilin-1 is downregulated in dHAND mutants. Neuropilin-1 was isolated as a dHAND-dependent gene by subtractive hybridization between dHAND-mutant and wild-type hearts (a). Quantitative RT-PCR of neuropilin-1 in wild-type (+/+) and dHAND-null (–/–) hearts confirmed downregulation of neuropilin-1 in dHAND-mutant heart and yolk sac RNA (b). Cycles of PCR amplification are shown. RNA loading was controlled by assaying GAPDH expression. Whole-mount in situ hybridization demonstrated neuropilin-1 expression in the heart (ht) and aorta (arrowheads) of an E9.0 embryo in right lateral view (c). At E9.5 (d), neuropilin-1 expression, viewed from the left lateral side, was seen in the aorta and aortic arch arteries (arrowheads), heart, pharyngeal arch (pa), limb bud (lb), and septum transversum (st). In dHAND mutants, neuropilin-1 was downregulated in the rostral aorta, pharyngeal arches, and heart (e). Expression in the caudal aorta (arrowhead), limb bud, and septum transversum was intact. Transverse sections through the rostral part of embryos in d and e revealed neuropilin-1 expression in the endothelial (e) and mesenchymal (m) cells of the wild-type aorta (f), but no expression of neuropilin-1 in the dHAND-null aorta (g). h, head.