Morpholino-mediated knockdown of katna1 function with a translation-blocking (katna1aug1) morpholino inhibits spinal motor axon outgrowth. Embryos were injected with 1.8 pmol of control (CoMO) (B), 0.9 pmol of katna1aug1 (C) or 1.8 pmol of katna1aug1 morpholino (D), fixed at 28 hpf, and then immunostained with znp-1. An uninjected control is shown in A. The effects of katna1 and spast translation-blocking morpholinos on spinal motor axon outgrowth are independent of p53 (E). Axons were measured in embryos injected with 1.0 pmol CoMO + 1.6 pmol p53-specific morpholino (p53MO) (n=20), 0.6 pmol spg4atg1 + 1.2 pmol p53MO (n=21), or 1.0 pmol katna1aug1 + 1.6 pmol p53MO (n=17). Statistical significance was determined using ANOVA with Bonferroni’s multiple comparison test. ***P<0.001. (F) Immunoblots for katna1 and tubulin demonstrating that katna1aug1-injected embryos have reduced levels of katna1 protein compared with CoMO-injected embryos. (G) Immunoblots for spastin and tubulin demonstrating that spg4atg1-injected embryos show reduced levels of spastin protein compared with CoMO-injected embryos. The right panel in F and G is a section from an equivalently loaded Coomassie-blue-stained gel to demonstrate relative loadings. Scale bars: 50 μm.
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