Fig. 2

WNK1/HSN2 knockdown in zebrafish using antisense morpholino oligonucleotides (AMO).

A) Knockdown embryos show no morphological phenotype, but reveal posterior lateral line defects (PLL) as observed under fluorescence with the 4-di-2-ASP vital dye when compared with non-injected WT embryos at 72hours post-fertilization. The knockdown embryo presented is a representative result obtained from MO-hsn2-SB52 injection. B) Each neuromast of the PLL observed with 4-di-2-ASP is assigned a score and totals for each fish is tabulated by condition and presented as a box plot, showing significantly lower scores for knockdown embryos when compared with WT. C) Two human sequence constructs of WNK1/HSN2 were designed for the rescue of the knockdown phenotype: a partial sequence containing exons 1-HSN2 and a complete construct containing exons 1–28 (but missing exons 11 and 12). D) 4-di-2-ASP score was assessed for embryos injected with MO-hsn2-SB32 and concentrations of 30, 50 or 75ng/μl of either partial or complete human WNK1/HSN2 constructs, revealing a partial rescue of the knockdown phenotype for embryos injected with 50 and 75ng/μl of the complete construct. Scale bar: 100μm.