Extended Data Figure 8: Recursive splicing regulates the alternative splicing of RS-exons.

a, Qiaxcel analysis and quantification of the splicing intermediates of indicated CADM2 splicing reporter products following transfection in SH-SY5Y cells. Primers used are indicated by red arrows in schematic, together with expected products and their sizes. b, RT–PCR analysis of the zebrafish cadm2a mRNA after in vivo injection of AON-2. Sequencing reveals RS-exon inclusion results in subsequent splicing to additional downstream cryptic elements before the second exon, explaining why RS-exon included product size is larger than expected. c, qRT–PCR analysis of exon–exon junctions surrounding the RS-site containing introns following AON-A1 mediated inhibition of RS-site use of the human CADM1 and ANK3 genes (n = 3, 1 experiment) or the zebrafish cadm2a gene (n = 7, 3 separate experiments). d, Splice site scores of reconstituted 5′ splice sites following first step of recursive splicing versus the 5′ splice sites of corresponding recursive exons.