Fig. S2

Splice site- and translation initiation-blocking morpholinos significantly decreased Nr2e3 protein levels but caused no overall morphological defects. (A) Representative images of non-injected controls (NICs) and nr2e3 splice morphants at the indicated stages. (B) Semi-quantitative RT-PCR demonstrates a band shift consistent with retention of the intron 2 of nr2e3 (88 bp in length), which is predicted to lead to a premature stop codon. (C) Representative images of antibody labeling of Nr2e3 of the pineal complex at 1 dpf in NICs and nr2e3 ATG morphants. Average fluorescence is 263,923 ± 36,837 arbitrary units in NICs (a.u., n=8) and 8,750 ± 4,063 a.u. in nr2e3 ATG morphants (n=7); P = 2.27 x 10^-5; two-tailed t-test. (D) Representative images of antibody labeled pineal complexes of NICs and nr2e3 splice site- morphants at 4 dpf. Quantification of all cell types in nr2e3 MO^splice can be found in Table 2. (E) Representative images of in situ hybridization of tbx2b in NICs and nr2e3 splice morphants at 24 hpf. Dorsal views. Scale bars: 30 µm.