Figure 5

Splicing effects of mutations in hMSH2 exons 6 and 10, and hMLH1 exon 17 in the three different cell lines. Panels A (hMSH2 exon 6), C (hMSH2 exon 10), E (hMLH1 exon 17) show the RT-PCR results of the splicing assay performed in the three cell lines transfected with 1 μg of the mutated and corresponding normal constructs. 48 hours after transfection, RNA was extracted, reverse transcribed and amplified with primers SD6 and SA2. The RT-PCR products were resolved on GeneGel Excel, stained with ethidium bromide and quantitated with an image analyzer (see methods). Mw = size standard. The percentage of exon inclusion is indicated below each lane. The graphs (B for hMSH2 exon 6 and its mutations, D for hMSH2 exon 10 and its mutations, F for hMLH1 exon 17 and its mutations) show the averages of percent exon inclusion (y-axis) in the three different cell lines when transfected with the constructs containing the different mutated or normal exons, representing the mean of two independent transfections done in triplicate for each construct (x-axis). Error bars represent standard deviation. The data were analysed using Student's t test (* = P < 0.05, ** = P < 0.01, *** = P < 0.001).