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Figure 3 Fluorescent whole-mount immunohistochemical detection of myoc in the yolk of zebrafish larvae (96hpf). Wild-type (A–C), heterozygous (D–F) and homozygous (G–I) myoc mutant embryos were incubated with 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 yolk. 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 on the yolk’s surface (arrows) (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 result observed in 10 embryos. The negative controls are shown in Figure S3D–F. Two-dimensional confocal image z-stacks are show in Supplementary Video S2.