Modulating Developmental Pathways in Zebrafish

All chemicals were dissolved in E3 medium and administered by immersion from 48 to 74 hpf. Vegf signalling was inhibited using 25 nM tivozanib (AV-951, AVEO Pharmaceuticals); vehicle control groups were exposed to 0.0025% dimethyl sulfoxide (DMSO). mTOR signalling was inhibited using 2-2.5 µM rapamycin (Sigma-Aldrich); vehicle control groups were exposed to 0.2-1% DMSO. MEK signalling was inhibited using 7.5-10 µM PD0325901 (Anastasaki et al., 2012 (link)) (Sigma-Aldrich); vehicle control groups were exposed to 0.75-1% DMSO. p38 MAPK signalling was inhibited by 25 µM SB 203580 (Sigma-Aldrich). Nos inhibition was achieved by incubation with 0.5 mM L-NAME (Sigma-Aldrich) diluted in E3 medium.