Fig. 1

Identification of the alternative splicing site within the mouse Ezh2 locus and the point mutation strategy. A Left panel shows alternative splicing sites and RT-PCR primer positions. Right panel shows RT-PCR result of the Ezh2 alternative splicing event in multiple mouse tissues. B Conservation analysis of the alternative splicing site sequence at mouse/human Ezh2 gene loci; the additional 27 nt DNA sequence in the long isoform encodes nine amino acids. The donor site sequence "GT" is marked in yellow. C WT mice expressed both the long isoform and the short isoform. Ezh2^Long expressed only the long isoform. Ezh2^Short expressed only the short isoform. Mutation strategy at Ezh2 alternative splicing site: mutation of the donor site sequence "GT" (the base marked in red is the sequence after mutation) to produce only the long isoform (Ezh2^Long) or the short isoform (Ezh2^Short) in mutant mice. D Sanger sequencing verified the base sequence of mouse point mutations. The arrows indicate the mutated bases in the mouse genome. E Representative images of genotyping PCR products visualized on agarose gels. F RT-PCR detected the expression of Ezh2 transcripts in testis and ovary of WT, Ezh2^Long and Ezh2^Short mice