Fus mRNA expression in early development of zebrafish and Fus levels are reduced upon KD with a specific AMO.
A) In situ hybridization with antisense (upper panels) and sense (lower panel) oligo probes specific to Fus mRNA revealed a predominant expression in the CNS, in particular in the hindbrain, spinal cord, as well as intersomitic segments as shown in B) in 24 hpf zebrafish embryos. C) Western blot analysis of zebrafish embryos injected with an AMO that specifically binds and inhibits Fus mRNA translation, showing reduced levels of Fus expression as compared to non-injected zebrafish embryos and zebrafish embryos injected with an AMO, where 5 nucleotides are mismatched (mismatch). D) WT human FUS mRNA or one of three ALS-related FUS mutations, each tagged with myc, were overexpressed in zebrafish and larval extracts were collected. Immunoblotting with myc antibody reveals similar expression in all these extracts.