Fig. 2

Minigene experiment. a The scheme shows the set-up of the minigene constructs for the splicing analysis in the WT and mutant expression vectors containing c.1200 + 1G > A mutation (arrows). b COS-7 cells were transfected with the wild-type (WT) or mutant (MT) minigene constructs. Total RNA from the transfected cells were used for RT-PCR of CYP11B1 cDNA. The figure shows the expected 560-bp PCR product from the WT construct and two shorter incorrectly spliced products from the mutant, sized 481 and 283 bp on an agarose gel. c Electropherograms of the minigene PCR products. The 481 bp mutant fragments skipped the entire exon 7, while containing the full-length sequences of exons 6, 8, 9. The 283 bp mutant PCR product skipped the exons 7 and 8, while retaining full-length sequences of exons 6 and 9. Black lines indicated exon–exon boundaries