Morpholino-mediated knockdown of the Jagged genes in zebrafish. (A-C) The jag1a splMO (A) and jag1b splMO (B) were designed to target the splice donor sites of the first exon-intron boundary. These splMOs caused deletions of the mRNAs by altering the splicing donor sites (*; confirmed by DNA sequence analysis of the PCR products indicated by yellow and red arrowheads in C). The altered splicing forms of mRNA potentially produce premature truncated proteins (A,B). Open and filled circles indicate the normal and cryptic splice donor sites, respectively. (C) RT-PCR analysis of Jagged gene expression in jag1a MO (jag1a splMO)- and jag1b MO (jag1b splMO)-injected embryos. Yellow and red arrowheads indicate the shorter forms of jag1a and jag1b cDNA, respectively. ef-1 was used as a reference gene. (D) The jag1b atgMO reduced the protein level of Jag1b (Jag1b). α-tubulin was used as a loading control.
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