Fig. 6: RT-PCR analysis of HeLa cells transfected with KCNQ1 c.1686−9 T > C minigene plasmids.

The agarose gel electrophoresis of minigene-derived KCNQ1 and the endogenous RPL19 RT-PCR products from two transfection experiments are shown in a and b using the indicated reference (REF) or mutant (MUT) KCNQ1 minigene plasmids (as described in Fig. 5). Amplification of KCNQ1 cDNA was performed with a forward primer located in E13 and a reverse primer in the vector-derived Myc epitope. Each RT-PCR lane (1–14) corresponds to a cell culture well processed independently. The amplification of RPL19 was used as a cDNA normalization control. NTR nontransfected cells, L DNA ladder. c DNA sequencing chromatograms of the longer 263-bp KCNQ1 band corresponding to the normal mRNA including exons 13–14–15 and the shorter 216-bp band with the complete skipping of exon 14 (ΔE14 KCNQ1). The deduced amino acid sequence is shown below the nucleotide sequence, as displayed by the Chromas software. The skipping of E14 introduces a frameshift (FS) at the start of E15. Full-size RT-PCR gel images are presented in Supplementary Fig. 5.