Fig. S1

Validation of Eaf1-MO2 and Eaf1-MO3. (A) Embryos co-injected with the standard morpholinoSTD and the MT reporter vector (mutated Eaf1-MO targeted region fused with GFP) or WT reporter vector (wildtype Eaf1-MO targeted region fused with GFP) demonstrated that STD could not block expression of the GFP fusion protein (a and b); embryos co-injected with Eaf1-MO2 and the MT reporter or WT reporter indicated that Eaf1-MO2 could not block expression of MT-GFP fusion protein (c), but could block expression of WT-GFP fusion protein effectively (d). (B) The schematic of eaf1 exon-intron structure, Eaf1-MO3 targeting position and RT-PCR primer locations. (C) (a) A 135-bp β-actin fragment was detected in all embryos by RT-PCR. (b) An expected 265-bp fragment resulted from normal splicing of eaf1 could be detected by RT-PCR in the wildtype embryos and the embryos injected with Eaf1-mismatch-MO (8ng/per embryo), but not in the embryos injected with Eaf1-MO3 (4ng/per embryo). (c) An expected 628-bp fragment resulted from splice-blocking of eaf1 could be detected by RT-PCR in the embryos injected with EAF1-MO3, but not in the wildtype embryos and the embryos injected with Eaf1-mismatch-MO. The embryos were collected at 9-10 hpf.