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Fig. 5
Phenotypic and quantitated effects of FICZ and αNF alone and in combination. Groups of 1 dpf embryos were exposed to different doses of FICZ or vehicle control (DMSO) in combination with αNF starting at 1 dpf. (A) and(B) Illustrative pictures of phenotypic effects; (A) control embryo (top) and embryo treated with FICZ (1.0 nM) + αNF (0.5 µM) (bottom). (B) Embryo treated with FICZ (0.005 nM) + αNF (1.0 µM). Abbreviations: reduced growth (RG), body curvature (BC), craniofacial deformities (CD), reduced eye size (RE), pericardial- and yolk sac edema (PE, YSE), and tube-shaped heart (TSH). All pictures were taken at 3 dpt (4 dpf). (C)-(F) quantitated data on lethality and phenotypic effects after exposure to (C) FICZ (0.005-10 nM; F0.005 etc), or (D) αNF (0.1 to 1 µM; A0.1 etc), or (E) combinations of αNF and FICZ. (F) Embryos injected with Ahr2-MO (Ahr) were exposed to FICZ (10 nM), αNF (1 µM), or a combination of FICZ and αNF. DMSO (D) was used as a vehicle control for all exposures. At 3 dpt (4 dpf) frequencies of cumulative mortality (Dead), pericardial- or yolk sac edema (Edema), and failure of swim-bladder inflation (-SB, no edema) were determined as% of total number of embryos at exposure start based on one representative experiment (n = 25-30). Embryos with none of the effects listed were scored as normal. Statistically significant differences among groups were determined by Fisher’s exact test with Bonferroni’s correction. A difference in mortality rate is indicated by asterisks (**p < 0.01; ***p < 0.001) between a treatment group and the reference group (D, A1, A0.5, or Ahr + D) and by daggers (†††p < 0.001) between an Ahr2-MO group and its corresponding non-injected group. For embryo groups where no significant mortality was observed, the difference in rates of any effect between an exposed group and the reference group is indicated by double daggers (‡p < 0.05; ‡‡‡p < 0.001).