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Fig. 10 a Notochord curvatures of variable severity (purple angles schematics) and bright filed images of WT and mutant ARF3 expressing fish at 30 hpf. The images are representative of embryos from three (WT, K127E, and L12V/D67V), two (P47S), and one batch (other mutations). b Rose diagrams showing notochord angles, no. of angles = 17 (WT); 49 (K127E); 32 (L12V/D67V); 31 (P47S); 35 (D93N) and 38 (T32N) pooled from a total n of embryos indicated below (c). Mean vector (µ) ± circular SD is shown. Dark and light violet shadings in the rose diagrams represent mild and severe classes of notochord curvatures, respectively. c Incidence of embryos with mild or severe notochord curvatures, set 1: no. of embryos = 9 (not injected); 17 (WT); 15 (K127E, ****p < 0.0001) and 11 (L12V/D67V, ***p = 0.0005) of three independent batches; set 2: no. of embryos = 13 (WT); 28 (P47S); 19 (D93N, ***p = 0.0003) and 24 (T32N, ****p < 0.0001) of one batch. Data are expressed as mean ± SEM (set 1) or mean (set 2). c’ Quantification of the number of notochord curvatures per embryo from one batch (same n of embryos as in c): K127E, ****p < 0.0001; L12V/D67V, **p = 0.0015; D93N, **p = 0.0022 and T32N, ****p < 0.0001. d Schematics of Krox20 and MyoD expression at 15 hpf. Black square brackets indicate AP and ML axes. R3 and R5: rhombomeres 3 and 5. e Bright-field images showing Krox20 and MyoD in situ mRNA staining (insets show severe cases). The images are representative of embryos from two independent batches for WT, K127E and L12V/D67V and from one batch for the other mutations. f Quantification of AP embryo extension, set 1: no. of embryos = 10 (not injected); 10 (WT); 10 (K127E, ****p < 0.0001); 10 (L12V/D67V, **p = 0.0078); set 2: no. of embryos 18 (WT); 22 (P47S, *p = 0.0169); 24 (D93N, *p = 0.0207) and 16 (T32N, ***p = 0.0002) of one batch. In c’ and f data are expressed as box-and-whisker with median (middle line), 25th–75th percentiles (box), and min-max values (whiskers). All the data points and the mean (“+”) are also shown. g Incidence of fish with different convergence and extension (CE) index values (same no. of embryos as in f) ****p < 0.0001 (K127E and L12V;D67V), **p = 0.0015; *p = 0.0251 (D93N), *p = 0.039 (T32N). Different datasets for the same measurement are shown in adjacent plots with the internal WT control for each set. Not injected controls between batches are not significantly different. Non-parametric Kruskal–Wallis followed by Dunn’s multiple comparison post hoc test (c’, f), Two-sided Chi-square’s test in 2 × 2 contingency table (c, g, normal vs. phenotype) are used to assess statistical significance. Source data are provided as a Source Data file.