Figure 3

Effect of Smc3 knockdown on embryogenesis. A) Western immunoblot analysis of Smc3 level in embryos injected at 1–2 cell stage with increasing doses of smc3-MO or smc3-mmMO. At 24 hpf 10 embryos were dechorionated and the yolk remnant removed. After homogenization in 100 μl lysis buffer, proteins were separated on 7.5% SDS-PAGE and Smc3 detected by immunoblotting using goat anti-human SMC3 antibody followed by ECL reaction. B) Morphogenesis in normal (a-e) and morphant (f-j) embryos injected with 8 ng of smc3-MO. At the indicated times embryos were collected and their morphology examined. Note the presence of necrotic cells in the eye and the cerebellar region of embryos at 15 hpf. At 24 hpf the majority of the morphants display head necrosis and a range of tail developmental defects including a reduction of the tail extension, the loss of chevron-like structure of the somites and the narrowing or disappearance of the notocord.