(a) Injection of 1 nl of a 0.25 mM solution of slc7a14-MO dramatically reduces eye size in zebrafish. (b) At 2dpf, 4dpf and 5dpf, the eye-to-body ratio of fish injected with the slc7a14-MO was significantly smaller than that of fish injected with the control MO. N=6, Dunnett test, ***P<0.001. (c) Light-induced locomotor response with sudden light-to-dark transitions demonstrated significant visual impairment in zebrafish injected with the slc7a14-MO. N=6 in each group. (d) Dunnett tests of the light-induced locomotor responses indicate that significant differences exist between the responses of fish with small eyes and those with normal eyes during the light-ON phase. N=10, Dunnett test, **P<0.01. Bars in the graph represent standard errors of the mean (s.e.m.).

Confirmation of MO injection in zebrafish Larvae. (a) After injection of fluoresceinated standard control MO and Slc7a14 MO solution, fluoresceins were observed in 1dpf Larvae. Scale bar, 500µm. (b) RT-PCR confirmed splicing change in Slc7a14-MO injected larvae. Three doses (1nL of 0.25nM, 0.15nM, 0.05nM) of MOs were tested. (c) Immunohistology demonstrated no significant changes of retinal structure in the Slc7a14-MO zebrafish. Each section was shown with optic nerve. Scale bar, 50µm.

Representative images of the MO-injected zebrafish. Comparing with the control MO-injected zebrafish, Slc7a14-knockdown led to dramatically smaller eye size. Scale bar, 1000µm.