Fig. 10

Morpholino-mediated knockdown of blt1 and lta4h affects epiboly.

(A) Diagram of a partial map of blt1 genomic DNA. Exons and introns are shown as boxes and lines, respectively. Two nonoverlapping blt1 MOs, blt1 MO and blt1 spl MO, were designed to target translation and splicing of Blt1, respectively. The blt1 MO should eliminate the transcription of exon 1, which contains the translation start site, resulting in aberrant protein synthesis. The efficacy of blt1 spl MO was validated by RT-PCR using blt1_fw and blt1_rv or blt1_rv2. Total RNA was isolated from blt1 spl MO or control MO-injected embryos at 24 hpf. Reduced expression of normal spliced blt1 transcripts and increased mis-spliced blt1 transcripts are shown. (B) Representative images of delayed epiboly of blt1 and lta4h morphants from 4 to 10 hpf. Embryos were injected with 2.5 ng MOs at the 1- to 2-cell stage. (C) Histogram illustrating the percentages of normal and delayed embryos. **, P < 0.005, one-way ANOVA with Bonferroni post-hoc test. Data are representative of at least two independent experiments with similar results.