Ocrl regulates receptor recycling and trafficking of endocytic ligands in zebrafish neuroepithelial cells. (A) Representative confocal images of endocytic uptake of RAP–Cy3 30 min post hindbrain ventricle injection in live ocrl mutant embryos (n=14) or WT controls (n=19). Scale bar: 10 µm. (B–E) Quantification of the average total fluorescence intensity per confocal slice (B), average number of RAP–Cy3 puncta per confocal slice (C), average RAP–Cy3 punctum fluorescence intensity (D) and RAP–Cy3 ventricle intensity (E) at 30 min post hindbrain injection of RAP–Cy3 in WT and ocrl mutant embryos. (F) Representative confocal microscopy images of fixed tissue sections from WT (n=17) or ocrl mutant (n=22) embryos stained with antibodies against EEA1. Scale bar: 5 µm. (G) Quantification of EEA1-positive endosome size in WT and ocrl mutant embryos. (H) Representative confocal microscopy images of fixed tissue sections through the zebrafish hindbrain from WT (n=14) or ocrl mutant (n=12) embryos stained with antibodies against Lrp2. Scale bar: 5 µm. (I) Quantification of the mean Lrp2 fluorescence intensity from WT or ocrl mutant embryos. a.u. arbitrary units. ns, not significant; *P<0.05; **P<0.01; ***P<0.001. Statistical comparisons between groups were made using two-tailed unpaired Student's t-test.
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