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Fig. s2 Validation of splicing assay. (A) specificity of primers used in RT-PCR splicing assay. Primers sets to identify Ex7 and Ex8 containing variants of both jnk1a and jnk1b transcripts were assessed against all 8 transcripts contained within plasmids, using primers for the plasmid backbone (M13) as loading control. Restriction enzyme digests showing original PCR product and complete digestion by enzyme to identify C-terminal extension in jnk1a and jnk1b transcripts. (B) Graphical representation of gel densitometry indicating the splicing assay was carried out during the linear phase of amplification for jnk1a and jnk1b Ex7 and Ex8 PCR (Arbitrary logarithmic Units). (C) PCR with plasmid containing transcripts were used as positive controls for PCR reactions and indicated efficiency of each reaction.