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Fig. 9 Apical polarity development in MZoep. ZO-1 staining in (A,B,C,D) wild-type and (E,F,G,H) MZoep embryos at different stages of neurulation. All images are maximum confocal projections of six consecutive z-levels taken from a dorsal view at posterior hindbrain and anterior spinal cord regions. Anterior is up. White arrows indicate the dorsal midline of the embryo. The ZO-1 staining surrounding the edge of the neural tissue is from the polarized enveloping layer overlying the neural tissue. Scale bar is 50 µm. (I) The average number of pixels of ZO-1 staining over time for MZoep and wild-type embryos. At 13.5 hpf the number of ZO-1 puncta in MZoep embryos is significantly different to wild-type (*P <0.01, two-way ANOVA), indicating that ZO-1 polarization is delayed by 30 minutes in MZoep embryos. At subsequent time points (14, 15 and 16 hpf), there is no significant difference between the wild-type and MZoep embryos. Error bars indicate the standard deviation. ANOVA, analysis of variance; hpf, hours post fertilization; MZoep, maternal-zygotic one-eyed pinhead; ZO-1, zonula occludens 1.