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Figure 4 Fluorescent whole-mount immunohistochemical detection of myoc in the tail of zebrafish embryos (96hpf). Wild-type (A–C), heterozygous (D–F) and homozygous (G–I) myoc mutant embryos were incubated with a chicken anti-myocilin (TNT) primary antibody and a Cy2-conjugated goat anti-chicken IgY secondary antibody. Three-dimensional reconstruction from z-stack scanned confocal microscopy images (A,D,G) of the tail. Sections (92 μm) 8 (B,E,H) and 16 (C,F,I) were selected from z-stack images to show the precise localisation of the green signal in the tail’s skin (arrow) (A–C). Blue: DAPI nuclear staining. Green: Cy2-conjugated goat anti-chicken IgY secondary antibody. Red: tissue autofluorescence. The cross indicates the position of the embryonic axes (D: dorsal; P: posterior; V: ventral; A: anterior). The images are representative of the results observed in ten embryos. The negative controls are shown in Figure S3G–I.