Fig 6. Alternative splicing analysis of Impad1 mRNA.

(A) RT-PCR using primers Imp7 and Imp12 that amplify a region spanning exon 1 to exon 5 was performed from skin total RNA of wild-type and Impad1^D175N/D175N newborn mice and analysed by 1.5% agarose gel. In wild-type animals one band, 639 bp long, corresponding to the correctly spliced Impad1 transcript including the 5 coding exons is present. This band is not detected in mutant mice, conversely two different bands, 477 bp and 383 bp long, respectively are observed. (B) Sequencing of the two bands demonstrates that the two transcript variants lack exon 2 or both exon 2 and exon 3.