Construction and expression analysis of the PCLO minigene.

(A) Representation of the PCLO minigene fused to the GFP coding sequence. The position of the mutation is indicated in bold. The positions of the primers used for RT-PCR are indicated by small arrows. (B) RT-PCR of cells transfected with the PCLO minigene constructs. The upper band denotes the unspliced transcript. The central band corresponds to a 676-bp spliced product. The lower band denotes a 575-bp alternative product. GAPDH was used as the control for normalization. (C) Splicing efficiency of the spliced and unspliced transcripts from three independent experiments. *Transcript splicing efficiency is the mean ratio of fluorescent intensity of correctly spliced transcripts: spliced plus unspliced transcripts for a given expression construct. Standard deviations from three independent experiments are shown. **Two-tailed P value calculated by Student’s t-test.