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. 2014 May 5;211(5):801–814. doi: 10.1084/jem.20131780

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© 2014 O’Donovan et al.

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Figure 1. — Conditional expression of kaB-RAF specifically activates MAP kinase signaling in the nervous system. (A) E18.5 mouse embryos expressing kaB-RAF develop grossly normally, except for substantial hydrocephalus. (B) DRG size is unaffected by kaB-RAF expression. (C) Representative Western blots show increased levels of pMEK1/2 and pERK1/2 in the E12.5 neocortex and spinal cord. (D) Quantitation of Western blots as in C. Amounts are normalized to corresponding βIII-tubulin levels. n = 4 repeats with three animals per group for each bar. Student’s t test: *, P < 0.05; **, P < 0.01. (E–G) Analysis of the interaction of kaB-RAF with other signaling pathways in the E12.5 nervous system. Representative Western blots are from three independent experiments, each with two or three E12.5 embryos for each genotype. (G) B-RAF protein levels are not increased in E12.5 DRG and spinal cord of LSL-kaBraf:nesCre embryos and control littermates. Molecular mass is indicated in kilodaltons. (H) Representative images of nerve endings in trunk skin of E13.5 embryos. Bars: (B) 200 µm; (H) 40 µm. (I) The number of branches per nerve trunk was quantitated, blinded to the genotypes, for a defined skin section by counting the total number of branch points observed along each major nerve trunk entering the skin (Zhong et al., 2007). n = 3 for each group. (D and I) Error bars indicate SEM.