PD0325901 is an effective MEK inhibitor in zebrafish development. (A) Chemical structure of the selective MEK inhibitor PD0325901. (B) Western blot of 10-hpf zebrafish embryos treated with increasing concentrations of PD0325901 from 4 hpf, and immunostained with anti-phospho-ERK1/2 and anti-ERK1/2 antibodies. (C) Images of developing zebrafish treated with DMSO, or 0.5 μM or 1.0 μM PD0325901. Zebrafish were imaged under brightfield lighting conditions (top row), or using fluorescence microscopy to visualise GFP expressed from the dusp6 promoter (dusp6-GFP; bottom row). GFP signal was first detected in gastrulating embryos at 50–60% epiboly (white arrowhead). The expression area expanded towards the vegetal pole during epiboly (white arrowhead) and at the tail-bud stage the expression was more diffused with higher expression levels at the tail bud (yellow arrowhead). By 24 hpf, GFP expression was detected in the brain (red arrowhead), along the spinal column and in the tail bud (yellow arrowhead). n=30 embryos per treatment condition.
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