|
Fig. S5
Recapitulation of phenotype by a ninl ex15 spMO (4 dpf).
(a-b) Injection of 4ng ninl ex15 spMO (n>100) results in heart edema and small eyes. No defects in body curvature were observed in comparison to control MO-injected larvae. (a', b') Analyses of bodipy-stained retinas of ninl ex15 spMO-injected larvae (n = 10) revealed defects in photoreceptor outer segment formation (10 of 10) similar to those observed in ninl atgMO-treated larvae, whereas stained retinas of control MO-injected larvae (n = 10) appeared normal (10 of 10). Scale bars represent 500μm (a-b) and 5μm (a'-b'). (c) RT-PCR analysis on RNA isolated from 25 larvae that were either uninjected, injected with control MO (6ng) or injected with various amounts of ninl ex15 spMO (2, 4, 6ng), collected at two different time points after injection (2 dpf and 4 dpf). One PCR product of the expected length (~500bp) was obtained from RNA from uninjected and control MO-injected larvae. Sequence analysis revealed that this was the predicted transcript including exons 13–16. RT-PCR analysis on RNA obtained from the morphant larvae resulted in two products: Sequence analysis of both fragments revealed that the shorter product is the predicted wild-type transcript (ex13-16) and that the longer transcript in addition includes the entire intron 14 (85 bp), resulting in premature termination of translation already after two codons in intron 14. This aberrant splicing persists at 4dpf.