Fig. S4

Fig. S4 Dose Dependency of Morpholino-Mediated Interference with U12-Type Splicing in Zebrafish Embryos
Embryos were injected with either a five-mismatch control morpholino (Co, at 0.1mM) or with increasing concentrations of an antisense U12 morpholino (0.002, 0.005, and 0.01 mM). Total RNA was prepared from 30-40 morpholino-injected embryos 18 hpf and splicing of U2-type (U2) and U12-type (U12) introns (in) was monitored by RT-PCR using primers hybridizing in the upstream and downstream exons, respectively. Amplification products were resolved by agarose gel electrophoresis and visualized by ethidium bromide staining. Splicing products are diagrammed on the right (exons, open boxes; introns, lines). mapk3, 8 and 12, mitogen-activated kinases 3 and 8, and 12, respectively; sms, spermine synthase. M, 100 base-pair (bp) DNA ladder, lowest band is 100 bp. -RT, corresponds to RT-PCR reaction lacking reverse transcriptase.