Flh acts in parallel with Tbx2b to specify parapineal fate, but does not govern migration. (A) Dorsal views of antibody labeling of parapineal neurons (krt4:eGFP+) as well as axons and dendrites (acetylated tubulin+) in the epithalamus at 5dpf. The pineal organ regions are marked by dashed circles. Scale bar: 30µm. (B) Quantification of the number of parapineal neurons in siblings (WT and flhn1+/-) or flhn1-/- mutants that were either non-injected (NIC) or injected with tbx2b morpholinos; mean±s.e.m. and number of samples (n) are shown. Loss of Tbx2b function suppresses the flhn1-/- supernumerary parapineal specification phenotype, as well as migration of parapineal neurons. ****P<0.0001, **P<0.01, P>0.05 (n.s.), one-way ANOVA with Tukey′s post-hoc analysis, comparisons with NIC;siblings (*), comparisons with tbx2b MO;siblings (#), and a comparison with flhn1-/- (+) are shown. (C) Quantification showing the percentages of larvae that display normal, bilateral, mixed (some neurons migrate and others do not), or medial (all neurons remain near the midline) parapineal-migration phenotypes and number of samples (n).
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