Fig. 3

BCS1L mutations in patient and parents. Sanger sequencing of the BCS1L gene in the patient, parents and control genomic DNA. a c.306A > T inherited from the father and (b) c.399delA inherited from the mother. c Sequencing of cDNA from the father showed the wild-type transcript and a small amount of the correctly spliced transcript carrying the c.306A > T mutation, whereas (d) sequencing of the mother’s cDNA shows expression of the transcript carrying the c.399delA mutation. e Transcript-specific RT-PCR analysis of the c.306A > T splice site mutation in patient and control fibroblasts. The upper gel shows a 346-bp fragment amplified from the patient (P) but not from the control (C) fibroblast cDNA, confirming the presence of incorrectly spliced transcript in the patient. The lower gel shows a 373-bp wild-type fragment amplified from both control and patients cDNA. Asterisk denotes a larger fragment likely from a partially spliced transcript retaining the 98-bp intron between exons 3 and 4. A fragment of similar size is also faintly detectable in the –RT (minus reverse transcriptase) control for the patient sample. H₂O denotes a control PCR reaction without template