FIGURE 2

FIGURE 2

Specific tuba8l2 knockdown in zebrafish induces axonal abnormalities. (a) An ATG morpholino was designed against the Danio rerio TUBA4A orthologue tuba8l2. (b–e) Western blot was performed at 48 hpf after injection of different doses of ATG morpholino against tuba8l2 (0.160 mM, 0.125 mM and 0.050 mM) as well as the injection of a control morpholino (0.160 mM). N = 3 experiments; n = 10–15 zebrafish per group per experiment. Quantification of tuba8l2 panel (c) and α-tubulin panel (e) protein levels relative to GAPDH for the different injection conditions. (f,g) Visualization of motor axons by SV2 immunohistochemistry at 30 hpf after injection of different doses of ATG morpholino against tuba8l2 (0.160 mM, 0.125 mM and 0.050 mM) or a control morpholino (0.160 mM). A non-injected condition was also included. Scale bar represents 50 μm p < 0.0001 (0.160 mM versus AMO control), p < 0.0001 (0.125 mM versus AMO control) and p < 0.0450 (0.050 mM versus AMO control); one-way ANOVA with Dunnett’s multiple comparisons. Axonal length was measured for N = 3 experiments; n = 10–15 zebrafish embryos per group per experiment; with every data point representing the average length of the five measured axons for each zebrafish embryo. *p < 0.05; ****p < 0.0001. AMO, morpholino; hpf, hours post fertilization.