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Fig. S7
Analysis of epistatic position of bsx function in pineal complex development.
(A-H,K-P) Dorsal views of embryos at 2 dpf after whole mount in situ hybridization. Anterior is to the top. Scale bar: 50 µm. (A,B) bsx expression is normal in embryos injected with otx5 morpholino (MO) (n=21/21) when compared to embryos injected with standard control MO (n=35/35). (C,D) otx5 MO function was confirmed through downregulation of irbp expression (n=22/24) in comparison to std ctrl MO injected embryos (n=34/34). (E) bsx is expressed in both pineal gland (PG) and parapineal (PP) in std ctrl MO injected embryos at (n=31/31). (F) In most of tbx2b MO injected embryos only PG bsx expression and no PP bsx expression was observed (n=41/43), in the other two embryos left-sided bsx expression was detected, but was weaker than in control embryos. (G) bsx expression is grossly normal in embryos injected with pitx2c MO (n=50/56). (H) bsx expression is also normal in embryos injected with nr2e3 MO (n=34/41). (I) Splice blocking activity of nr2e3 MO was confirmed through PCR (see Material and Methods section for details). (J) bsx expression in pineal complex of morphologically normal siblings in fgf8ati282a heterozygous incross (n=31/33). (K) No PP bsx expression is observed in embryos homozygous for the fgf8ati282a allele (n=10/10). (L) opn1sw1 is not expressed in the PG of std ctrl MO injected embryos (n=14/14). (M) otx5 knockdown by MO does not result in PG expression of opn1sw1 (n=26/26). (N) Heterozygous or wildtype siblings of bsx mutants do not express opn1sw1 in the PG (n=32/32). (O) Strong PG opn1sw1 is observed in all bsx mutant embryos (n=12/12).