Targeted SU5416 Treatment of Zebrafish Angiogenesis

SU5416 (SIGMA #S8442) was prepared and used as in Covassin et al. (2006) (link) and Stahlhut et al. (2012) (link). Briefly, a 200 μM SU5416 stock solution in DMSO (SIGMA #D8418; vehicle) was dissolved in fish water to a final concentration of 0.2 μM SU5416 (a suboptimal dose) and 0.1% DMSO. Control, vehicle-only (0.1% DMSO) treatments were also performed. Homozygous WT and homozygous plxnd1^skt6 mutant embryos were manually dechorionated before receiving the SU5416 and DMSO treatments using a common solution for both genotypes. Embryos were treated from 18 to 32 hpf to specifically target both Se and DLAV angiogenesis (see Torres-Vázquez et al., 2004 (link); Zygmunt et al., 2011 (link); Childs et al., 2002 (link); Isogai et al., 2003 (link); Zygmunt et al., 2012 (link); Yokota et al., 2015 (link)), and then fixed for immunostaining.