A Novel Alu Element Insertion in ATM Induces Exon Skipping in Suspected HBOC Patients

Figure 2

Mutant transcript missing ATM exon 54 identified and semi-quantified in patient-derived RNA. (a) Gel electrophoresis RT-PCR products from patients 1, 2, and 7 (P1, P2, and P7) and two control individuals (C1-2) using a forward primer located in ATM exon 53 and a reverse primer in ATM exon 55. The exon 54-skipped mutant transcript (Δex54) of 267 bp next to the full-length (FL) transcript of 350 bp is present in ATM insAluYa5-carriers and absent in controls. (b) Sanger sequencing electropherogram of the mutant transcripts across the exon 53-exon 55 junction. (c) Capillary electrophoresis of RT-PCR products of patient 1 (P1) and one control sample (C1) depicting the full-length (FL) and mutant (Δex54) transcripts in the patient, and only FL transcript in the control. (d) Capillary electrophoresis results are shown as bar diagrams depicting the percentage of full-length (FL) and mutant (Δex54) transcripts in patients 1, 2, and 7 (P1, P2, and P7) and two control individuals (C1-2) next to each other. Nonsense-mediated decay was inhibited by cycloheximide treatment. Data are represented as of three independent experiments. Bp: basepairs; Ex: exon; nt: nucleotides; NTC: no template control; NRT: no reversetranscriptase control.

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