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Fig. 2 Effect of genetic attenuation of tpcn2 on the number and shape of notochord vacuoles in embryos at ∼24 hpf. (A) Bright-field image of an intact wild-type zebrafish embryo at ∼24 hpf and (B) schematic showing the basic structure of the notochord from a lateral view. In (A), the black rectangle indicates the location of the notochord (adjacent to somites 8–11), where higher magnification images were acquired. (Ca-d) Representative bright-field images showing lateral views of the notochord at higher magnification in a (Ca) wild-type (ABTU; n = 5) embryo, (Cb) tpcn2 homozygous mutant (tpcn2−/−; n = 7), (Cc) p53-MO-injected embryo (n = 6) and (Cd) tpcn2-T-MO + p53-MO-injected embryo (n = 6). In these images the black dashed lines indicate the dorsal and ventral boundaries of the notochord. Scale bars, 200 µm (A) and 20 µm (C). (D) Violin-plots (indicating the mean ± SEM) onto which are superimposed individual data points showing the (Da) number and (Db) circularity of vacuoles in the control, morphant, and mutant embryos shown in (C). An explanation of what these parameters refer to is provided in the Experimental Procedures. The data were compared using one-way ANOVA and Fisher's least significant difference (LSD). Significance is indicated as *p < .05, and ***p < .001.