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Fig. 4. Knockdown of Nemp1 by MOab disrupts eye development. (A) MOab inhibits splicing of nemp1 pre-mRNA. (Upper panel) Sequence alignment of exon 1–intron 1 splice junctions. Target sequences of MOa and MOb are underlined. (Lower panel) RT-PCR detection of unspliced transcripts in MOab (25 or 50 ng) or 5mmMO (50 ng)-injected embryos. RT−, RT-PCR without reverse transcriptase reaction. (B) Morphological appearances of injected embryos. MOab (Ba), 5mmMO (Bb), or MOab plus nemp1 mRNA (Bc) were injected together with nβ-gal mRNA as a tracer (red) into the animal pole region of both right blastomeres at the four cell stage. Injected doses (per embryo): MOab 20 ng; nemp1 mRNA, 200 pg. Upper panels, dorsal view (anterior is upwards); lower panels, right side view of the same embryo in the upper panels. (C) WISH analysis of 5mmMO or MOab-injected embryos at stages 13–14. Arrowhead, injected side. |
