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. 2013 Jan 4;8(1):e53490. doi: 10.1371/journal.pone.0053490

Figure 3. Identification of exon skipping in mRNA from lag/lag mouse.

Figure 3

(A) Northern blot analysis of Kif14 mRNA. PolyA RNA from whole brains of +/+, lag/+, lag/lag mice was subjected to agarose gel-electrophoresis and then transferred to a nylon membrane. The membrane was hybridized with a Kif14 cDNA probe. (B) Transcripts analysis of Kif14 splice acceptor site mutation. Agarose gel-electrophoresis of Kif14 PCR products from first-strand cDNA prepared from whole brains of +/+, lag/+, lag/lag mice. (C) Sequence analysis of the three Kif14 transcripts identified in the lag/lag mouse. The Kif14 transcript in the lag/lag mouse exhibited skipping of an 11-bp segment of exon 5, entire exon 5, and exons 5 and 6 as a result of a G to A substitution at the acceptor site of exon 5. (D) Western blot analysis of Kif14 protein at E14.5 and P12. Whole brain extracts (40 µg of proteins) from +/+, lag/+, lag/lag mice at E14.5 and P12 were subjected to SDS-PAGE, followed by immunoblotting with the anti-Kif14 rabbit polyclonal antibody. Arrow indicates full-length Kif14. Asterisk indicates the non-specific band.