Fig. 6

The expression levels of anf and cmlc2 were altered in the zlrrc10 morphants. (A) CMO (a, c, e, g, i and k) and MO1 (b, d, f, h, j and l) injected zebrafish embryos at 2 dpf were subjected to whole-mount in situ hybridization using digoxigenin labeled anf (a–d) and cmlc2 (e–h) antisense riboprobes. Images of whole-mount immunostaining using MF20 antibody are shown in k and l. i and j are the same fields as k and l, respectively. c, d, g and h are ventral views and others are lateral views, all with anterior to the left. Note the marked increase in anf expression in the zlrrc10 morphant (b and d) but the reduction of cmlc2 expression in the morphant (f and h). A ventral view of the morphant clearly shows the small ventricle and a looping defect (h). (B) For quantitative analyses of gene expression, RT-PCR was performed on anf and cmlc2. β-actin was used as a control.