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Fig. 6. Sipa1l3 and Epha4 influence early eye specification. (A,C) WMISH at stage 13 revealed that Sipa1l3 (A) or Epha4 (C) function is required for proper rax and pax6 expression whereas sox3 is not affected. Red arrowheads indicate reduced marker gene expression at the injected side. Scale bars: 1000 µm. (B,D) Quantification of the data in A,C. (E,G) Knockdown of Sipa1l3 (E) and Epha4 (G) resulted in significantly reduced rax and pax6 expression domains (red arrowheads) compared with internal control as well as control MO-injected embryos at stage 23. Scale bars: 500 µm in overview; 250 µm in close-up views. (F,H) Quantification of the data in E,G. (I) rax RNA restores the Sipa1l3 MO-induced ocular phenotype. Red circles indicate eye areas. Scale bars: 500 µm in dorsal, lateral views; 250 µm in detail views. (J) Quantification of the data in I. (K) rax RNA rescues the Sipa1l3 MO-induced microphthalmia phenotype. (L) rax RNA restores the Epha4 MO-induced ocular phenotype. Red circles indicate eye areas. Scale bars: 500 µm in dorsal, lateral views; 250 µm in detail views. (M) Quantification of the data in L. (N) rax RNA rescues the Epha4 MO-induced microphthalmia phenotype. N, number of analyzed embryos in total; n, number of independent experiments. Error bars indicate standard error of the means (s.e.m.); *P≤0.05; **P≤0.01. P-values were calculated by a nonparametric, one-tailed Mann–Whitney rank sum test. |
