Fig. 4. Exon definition occurs in yeast.
(a) A plasmid containing the WT DYN2 gene or various mutants was transformed into a DYN2 KO strain. The splicing efficiency of intron 1 and 2 were evaluated using qRT-PCR with primers specific for intron 1 or intron 2 (indicated by arrows in the schematics under the bar diagram) normalized to total mRNA. Dots represent three technical replicates. (b) RT-PCR of RNA extracted from yeast strain carrying indicated plasmids, using primers located in exons 1 and 3 of Dyn2. A schematic of the splicing product and their expected sizes are shown on the right side of the gel. RT-PCR products using primers in exon 3 (bottom gel) serve as an internal quality control of the samples. Experiments in Fig. 4 were repeated two additional times with similar results.