|
Fig. 2
Efficiency and specificity of oep ASO-induced oep (one-eyed-pinhead) mutant phenotypes.
A) Injection of oep ASO induces dosage-dependent oep phenotypes that resemble zygotic (Zoep) and maternal-zygotic (MZoep) oep genetic mutants. B) oep ASO and MALAT1 ASO knockdowns are specific. The RNA levels of oep and MALAT1 were measured by qPCR in oep ASO (100 pg) and MALAT1 ASO (80 pg)-injected embryos. Shown is the fold change in RNA level compared to WT (wildtype), normalized to ef1a (error bars: standard deviation of the mean of 3 independent experiments). C) qPCR-based measurement of oep RNA levels in individual oep ASO (100 pg)-injected (red) or uninjected (black) embryos at shield stage (6 hpf). D) Rescue of oep ASO-induced oep phenotypes by coinjection of an oep ASO-resistant RFP-oep fusion mRNA. Note that the oep ASO-sensitive RFP-oep fusion mRNA is efficiently knocked down (no red fluorescence) and does not rescue. E) Quantitation of survival at 24 hpf and F) quantitation of phenotypic strength in survivors at 24 hpf in the presence versus absence of p53 (p53 MO-injected embryos) or RFP-oep fusion mRNA rescue construct. The number of embryos in each category is indicated.