Fig. S4

Fig. S4 MO-BNip1 rescues photoreceptors but not defects in OS and intracellular cisternae formation, related to Figure 3
(A) Zpr1 (green) and anti-rhodopsin (magenta) antibody labeling of 84 hpf wild-type and coa mutant retinas injected with MO-BNip1a, MO-BNip1b and 5mis-MO-BNip1a. Zpr1-positive photoreceptors are partially rescued in coa mutant retinas injected with MO-BNip1a and MO-BNip1b (arrowheads). (B) The percentage of zpr1-positive area relative to the total retinal area. Green and black bars indicate the means and standard deviations, respectively. Numbers of retinal sections used and p-values of t-tests are shown in Table S2. **p<0.01. (C) Western blotting of the heads of zebrafish 48 and 84 hpf non-injection embryos, and embryos injected with MO-BNip1a, MO-BNip1b and MO-sec22ba using anti-BNip1a, anti-BNip1b and anti-sec22L1 antibodies. Bands corresponding to BNip1a, BNip1b and Sec22ba were specifically reduced in bnip1a, bnip1b and sec22ba morphants, respectively (red arrow). Histone H3 is a loading control detected on the same membrane. (D–F) EM images of coa mutant photoreceptors injected with both MO-BNip1a and MO-BNip1b at 84 hpf. (E, F) Higher magnification. The OS is still small. The mitochondria (arrows, D–F) accumulated but tubular cisternae were reduced and abnormally positioned between the OS and the nucleus (n) (arrowheads, E). Some mitochondria were abnormally located near the nucleus (black arrows, E). Excessive accumulation of vesicle-like structures was observed (asterisks, F). The arrowhead indicates a connecting cilium (F). Abbreviations: n, nucleus; OS, outer segment. Scale bars: 50 μm (B), 0.5 μm (D–F).