Figure 8.

Crb2a localized to the apical surface of certain regions of the diencephalon neuroepithelium in nok^m520 mutants but not in nok^m227 mutants at 36 hpf. A, B. Arrows indicate the local enrichment of Crb2a at the apical surfaces of certain regions of the diencephalon in nok^m520 mutants. A boxed region in panel A is enlarged in panel B. C, D. Arrowheads indicate the corresponding apical surfaces of the diencephalon in nok^m227mutants, showing the lack of apical enrichment of Crb2a. A boxed region in panel C is enlarged in panel D. The apical markers ZO-1 localized ectopically to the interior of the retina (double arrows), as expected in nok mutants (A, C). The nuclei were stained with TO-PRO (green). Scale bar, 50 ! m. E. Western blotting analyses of protein expression at 30 hpf detected Crb1 in both nok^m520 and crb2a^m289 mutants as well as Crb2a in WT and nok^m520 but not in crb2a^m289 mutants. ! -tubulin blotting was used as loading controls. Please note that even though there are variations in ! -tubulin loading control signals for the Crb2a blotting (likely due to sample lose during preparation), our conclusion that certain level of Crb2a is present in nok^m520 mutant embryos still stands.