Fig. 1: RNA splicing analyses.

A Structure of the PMS2 gene and relative position of the four variants analyzed in this study. B Minigene splicing assays. RT-PCR primers are represented by black arrows (5’FAM modification indicated by *). The graphs show the average of 3 independent experiments. C Analysis of RNA extracted from the peripheral blood of a patient heterozygous for PMS2 c.1004A>G. Left panel, RT-PCR results obtained with RNA from the patient analyzed in parallel of equivalent samples from three control individuals (C1, C2 and C3). The graph shows the average of 2 independent experiments. Right panel, results from allele-specific expression (ASE) analysis using the SNaPshot primer extension method. The position of the primer used in the extension reaction is shown by the grey arrow. The fluorochrome electropherogram images show the outcome of 1 out of 2 experiments with similar results. The identity of the peaks (G and A) was converted to reflect the sequence in the sense strand. The ASE value indicates the level of expression of the variant allele (G) relative to the WT allele (A) calculated by normalizing the fluorescence obtained with the cDNA template with that obtained with gDNA. WT, wild-type.