Fig. S4

Fig. S4 Validation of the zebrafish-specific Rad21 antibody. Protein lysates were prepared from 20 wild type (WT), sibling (sibs) and rad21^nz171 homozygous mutant (mts) embryos at 27 h.p.f. A, Immunoblot showing Rad21 protein. B, α-tubulin loading control. Although there is less total protein in the mutant sample, the Rad21 protein is considerably more reduced in rad21^nz171 mutants than can be expected due to loading discrepancy alone. As expected, siblings, comprising a mix of wild type and rad21^nz171 heterozygotes, show intermediate levels of Rad21 protein. The band pattern detected, with the major band above 130kDa, is similar to that detected with commercial anti-human Rad21 antibodies.