Fig. 1

Phenotypes of atp6v1b2 zebrafish morphants. (a) The zebrafish atp6v1b2 gene was targeted by specific MO antisense to prevent proper splicing of exon 4 (E4I4-MO). Primers 2F and 5R interrogate the presence of wild type (non-mutant) transcripts or those in which intron4–5 has been inserted. Left: RT-PCR of atp6v1b2 transcript from control-MO and E4I4-MO injected embryos 2 days after fertilization, demonstrating insertion of intron4–5. Right: Sanger sequencing of both the wild type band and the intron4–5 inserted band validating the wild type sequence and the intron4–5 inserted sequence. (b-e) Compared to WT embryos (n = 10 per group), atp6v1b2knockdown group showed decreased body length (b, black dotted line, 32 hpf), brain ventricle size (b, green area, 32 hpf), severe pericardial oedema (b, black arrow, 32 hpf), inhibited inflation of the swim bladder (c, red arrow, 5 dpf), shorter pectoral fins (d, blue bracket, 6 dpf), and decreased hair cells (e, white star target, 5 dpf). Scale bars represent 100 μm. (hpf, hours post fertilization; dpf, days post fertilization). (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)