Figure 5

Downregulation of lpgat1 induced various embryonic developmental defects using morpholino oligonucleotides (MO). Each of embryos injected with MOs were observed overtime by time-lapse imaging. (A) The representative images of the normal embryos and larva, coagulated eggs, embryos and larva with poor-growth/pigmentation defect, and malformed embryo (with bent body axis, short body length and/or edema). LAS software (Ver. 4.12, Leica) was used to process the images. hpf; hours post fertilization. Scale bar, 500 µm. (B) Upper graphs, percentage of coagulated eggs after injection of morpholino antisense oligo (MO) specific to lpgat1 (MO1) or control MO (cMO) in combination with lpgat1 mRNA. MO1 is a splicing inhibitor, which bind to the exon–intron junction of immature lpgat1 mRNA and inhibit the splicing of pre-mRNA. Middle graphs, percentage of pigmentation defect and bottom graphs, malformation in uncoagulated embryos. Basically, the same results were obtained as shown in Fig. S9B using morpholino oligonucleotides 2 (MO2). The means of the data from three independent experiments for each group with n = 200 in total are shown. Error bars are S.D. Statistically significant differences are marked with asterisk. *p < 0.05; **p < 0.01, ***p < 0.001. Two-way ANOVA, Holm’s multiple comparison test was used.