Fig. S1

Knock-down of atp6v0c2 in zebrafish embryos. A: Schematic diagrams of atp6v0c2 genomic DNA and splice sites targeted by E2I2 MOs (atp6v0c2 MO). atp6v0c2 MOs block the excision of the intron between exon 2 and exon 3, generating abnormal atp6v0c2 mRNA in which exon 2 and exon 3 are separated by the second intron (I2). B: Confirmation of the efficiency of atp6v0c2 MOs by reverse transcriptase-PCR (RT-PCR) analysis. RT-PCR with RNA from control embryos amplifies a 677-bp segment of the transcript. However, RT-PCR using RNA from atp6v0c2 MO-injected embryos failed to generate a PCR product, because excision of the 2,169-bp intron (I2) between exon 2 and exon 3 did not occur. C: Confirmation of the efficiency of atp6v0c2 MOs by in situ hybridization with atp6v0c2 RNA probe. All panels show lateral views of wild-type and atp6v0c2 MO-injected embryos analyzed by atp6v0c2. Since atp6v0c2 MOs interfere with splicing between exon2 and exon3 and generate an mRNA split by intron2, whole-mount RNA in situ hybridization did not detect any atp6v0c2 expression in the embryos injected with atp6v0c2 MOs.