Fig. 3

Continuous suboptimal PD0325901 treatment prevents zebrafish BRAF^CFC phenotypes. (A) Wild-type embryos were injected with human BRAF^WT, the kinase-activating and most common CFC allele BRAF^Q257R, or the kinase-inactivating BRAF^G596V, and imaged at the indicated developmental time post-fertilisation. (B) Images of untreated control zebrafish (top panel) and a zebrafish expressing the kinase-inactivating BRAF^G596V allele under the melanocyte-specific mitfa promoter (bottom panel). Insert image depicts a high-magnification image of ectopic melanisation and nevi formation. (C) Continuous treatment of BRAF^WT- and BRAF^CFC-injected embryos with a suboptimal dose (0.2 μM) of PD0325901 from 4 hpf through 4 dpf. (D) Western blot of 10-hpf wild-type [uninjected (Un)] or BRAF^CFC embryos treated with 0.2 μM of PD0325901 from 4 hpf, and immunostained with anti-phospho-ERK1/2 and anti-ERK1/2 antibodies.