RT-PCR analysis of GFP splice variants in prp18a-1 mutants. RT-PCR analysis of GFP splice variants in prp18a-1 mutants. (A) Semi-quantitative RT-PCR was used to assess the accumulation of unspliced GFP transcript and two splice variants (resulting from splicing the canonical GT-AG and non-canonical AT-AC introns, respectively) in triplicate samples of the prp18a-1 mutant and the wild-type T line. Actin is shown as a constitutively expressed control. RT- and RT+ panels show experiments with and without reverse transcriptase, respectively. gDNA (T), genomic DNA isolated from T line. (B) The percentages of three major GFP RNA splice variants as determined from an analysis of RNA-seq data (Table S2). The average of five biological replicates is shown. The amount of total GFP transcripts did not change significantly in prp18a-1 mutants. Tatsuo Kanno et al. G3 2018;8:1367-1377 ©2018 by Genetics Society of America