(A) Quantitative PCR analysis of indicated genes expression using dissected eyes at stage 38. Quantification of normalized fold expression of indicated genes with one-way ANOVA (Dunnett’s multiple comparisons test), ****, P < 0.0001. Histograms represent means ± s.d. ns: no statistical differences between the groups.
(B) Inhibition of Hh pathway causes eye pigmentation loss. Eye diameter was quantified with unpaired t test, scale bar, 400 μm. Scatterplots represent means ± s.d from three biological repeats, *P < 0.05. Percentage of embryos with eye pigmentation loss was quantified with unpaired t test. Histograms represent means ± s.d from three biological repeats, **, P<0.01.
(C) Embryos were treated with DMSO or cyclopamine from stage 18–19 and then sectioned and immunostained with indicated antibodies at stage 39. Quantification of relative RPE65 fluorescence intensity with unpaired t test, scale bar, 40 μm. Scatterplots represent means ± s.d from three biological repeats, ****P < 0.0001. Quantification of relative rhodopsin fluorescence intensity with unpaired t test, scale bars, 40 μm. Scatterplots represent means ± s.d from three biological repeats, ****P < 0.0001.