Fig. 1

Egln2 downregulation rescues motor axonopathy in SOD1G93A zebrafish (A) Schematic representation of the splice-block antisense oligonucleotide morpholino (AMO) that targets the exon 2-intron 2 (E2I2) junction in zebrafish egln2 pre-mRNA, resulting in a differentially spliced egln2. (B) Validation of the AMO-induced splice defect using RT-PCR. Wild-type (WT) egln2 cDNA in 30 h post-fertilization (hpf) non-injected (NI) and control (ctrl) AMO or alternative splice product induced by E2I2 splice-blocking AMO, indicated by the arrowhead. (C) Representative images of synaptic vesicle glycoprotein 2a (SV2) staining at 30 hpf in zebrafish injected with SOD1WT RNA, SOD1G93A RNA, and SOD1G93A RNA with egln2 AMO. Scale bar, 50 μm. (D) Quantification of the axonal length. (E) Quantification of the percentage of abnormally branched axons. Data represent mean ± SEM with individual values shown (N = 8 experiments, each with 15 zebrafish per group). Statistical analyses were performed in (D) by a one-way ANOVA with Sidak’s multiple comparison test and in (E) with the Kruskal-Wallis test with Dunn’s multiple comparison test (∗p < 0.05, ∗∗∗p < 0.001, and ∗∗∗∗p < 0.0001).