Figure 4.

Noncanonical function of Hars represses vegfa expression. (A–D) In situ hybridization reveals that the expression of vegfa in the head is upregulated in about 25% of embryos at 26 and 36 hpf (arrowheads). (E) Quantitative real‐time PCR shows that vegfa mRNA levels in hars ^−/− are significantly upregulated at 48 and 72 hpf. Target transcript levels were normalized to the reference gene elf1a and to levels in WT controls, which were set to 1; n = 3 pooled biological replicates of ≥ 20 embryos, with three technical replicates per n. (mean ± standard error of the mean, *P < 0.05, two‐tailed t‐test). (F–K) SU5416 treatment suppresses ectopic branching in the brain and trunk of hars ^−/− at 60 hpf. Scale bars, 50 μm.