Transcript levels of pineal gland markers are greatly reduced in adult bsx−/− mutant zebrafish. Analyses of transcript levels of bsx (A) aanat2 (B), agrp2 (C), exorh (D), otx5 (E), and crx (F) at daytime (ZT6) and nighttime (ZT18) as determined by qRT-PCR. Copy numbers were normalized to the geometric means of copy numbers of eef1a1 and actb2, which were detected at high and stable levels in all samples; copy numbers were further divided by 100 to account for abundant transcript levels. Two-way ANOVA detected significant effects of time of day on transcript levels of aanat2 (p < .0001) and significant effects of genotype on transcript levels of bsx (p < .0001), aanat2 (p < .0001), agrp2 (p < .0001), exorh (p < .0001), otx5 (p < .001), and crx (p < .001). Transcript levels of homozygous (bsx−/−) and heterozygous (bsx+/−) mutants were compared to those of wildtype (bsx+/+) controls. Values on the graphs represent mean± SEM; n = 4, where each sample is pooled tissue from three animals. p-values as determined by multiplicity-adjusted Dunnett's multiple comparisons tests: ns, not significant, p > .05; *, p < .05; **, p < .01; ***, p < .001; ****, p < .0001. ZT, Zeitgeber time
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