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Fig. 1
Morphological analysis of zebrafish lacking Mcm2. (A) Autoradiogram to show the binding efficiency of the Mcm2 MO. In vitrotranslation of Mcm2 was done in the presence of CTRL (CTRL MO) and translation blocking MO (Mcm2 MO) using a cell-free reticulocyte lysate and 35S-labeled methionine. As template, pCS2+ containing parts of the 5′-UTR fused to the ORF of Mcm2 was used. n = 3 MO binding tests (in triplicate). (B) RT-PCR of non-injected (NI), control injected (splCTRL) and splice blocking injected (splMO) embryos at 24 hpf. In embryos with Mcm2 splMO the original band at 322 bp partially disappeared. Instead a second band at 1685 bp could be detected, which contained parts of intron 2. n = 2 independent experiments. (C) Live images of zebrafish at 48 hpf. Scale bars: 500 μm. (D) Mcm2 depleted embryos develop smaller anterior structures. Numbers of embryos analysed are given below the bars. 3–10 independent experiments. **** indicates a P value ≤0.0001, *** means P≤ 0.001. One-way ANOVA with Sidak's multiple comparison test. (E) Loss of Mcm2 impairs eye development. Numbers of embryos analysed are given below the bars. 3–6 independent experiments. **** indicates a P value ≤0.0001, ** means P ≤ 0.01. One-way ANOVA with Sidak's multiple comparison test. (F) Zebrafish lacking Mcm2 show a tendency to develop pericardial edema. Numbers of embryos analysed are given below the bars. 3–11 independent experiments. * indicates a Pvalue <0.05. One-way ANOVA with Sidak's multiple comparison test.