Table II. Epiboly percentage in ethanol and retinoic acid treated embryos.
Embryos were treated from 3 hpf until 8 hpf (during gastrulation) with 100 mM ethanol, 10−9 M retinoic acid, or 100 mM ethanol and 10−9 M retinoic acid together, and compared to untreated embryos (normal embryo medium: Control). At 8 hpf embryos were fixed using paraformaldehyde to stop development, and these embryos were imaged using a stereo microscope. The percentage of the yolk cell that is covered by the embryo proper was measured using ImageJ image analysis software (see Materials and Methods). Two-way ANOVA was performed to compare treatment groups. Ethanol-by-retinoic acid interaction was significant (P=0.0001), and pairwise comparison P-values are shown. Bonferroni adjustment for multiple (six) comparisons was used to ensure an overall ninety five percent confidence. Thus, P-value≤0.008 was used to conclude that groups were statistically different.
| Control | Ethanol | Ethanol+RA | RA only | |
|---|---|---|---|---|
| Mean | 85.4 | 74.0 | 78.1 | 79.8 |
| SD | 4.2 | 5.7 | 7.5 | 7.5 |
| n | 29 | 28 | 29 | 29 |
| P (vs Control) | <0.0001 | <0.0001 | 0.0012 | |
| P (vs Ethanol) | 0.0163 | 0.0009 | ||
| P (vs RA) | 0.3268 |