Fig. S2

brca2transcript analysis inzepmutants andbrca2morphants, and live morphology ofbrca2morphants. (A) RT-PCR analysis of brca2 transcripts in pools of zep embryos compared to WT control siblings (which included WT and heterozygous embryos), where embryos were scored into these groups based on edema at embryonic day 6. WT/heterozygous pools had normally spliced transcripts as well as variants that corresponded to inclusion of the 92 basepair intron 20–21 (see band doublet). zep mutant pools contained only the longer variant with the intron (top band). (B) RT-PCR of brca2 transcripts in brca2 MO2 injected embryos compared to WT control siblings revealed inclusion of the 92 basepair intron 20–21 only in the morphant pool, and WT transcripts were not detected. (C) WT embryos were microinjected with a brca2 start morpholino (MO1) or a morpholino designed to the splice acceptor site (MO2) of exon 21, and live morphology was observed over the subsequent 6 days of development. Morphants recapitulated the zep mutant embryo phenotype such that they developed pericardial edema (black arrowhead), starting by 72 hpf, which progressed in severity through the 144 hpf stage. Embryos are shown in lateral views.