Fig. 2

Molecular findings a Pedigree of the family described. Black arrow indicates index patient. Black symbols indicate affected subject. E: epistaxis, p/c AVMs: pulmonary/cerebral arteriovenous malformations, T: telangiectasias, cDM: cortical development malformation. Genotypes of available subjects are indicated below each symbol. b Molecular studies performed on genomic (gDNA) and complementary (cDNA) DNA in affected subjects. cDNA was obtained from blood-extracted RNA (Tempus Blood RNA isolation kit, Life Technologies) by retrotranscription (Maxima cDNA Synthesis Master Mix, Life Technologies). PCR and RT-PCR amplicons were analysed by direct sequencing: electropherograms show the nucleotide change c.3G > A in exon 1 of ENG. c Quantitative RT-PCR analysis of ENG transcript levels (probe: Hs.PT.58.4962347, IDT) in blood-extracted RNA from affected subjects of the study (ENG MUTATED, n = 3) and healthy subjects (n = 6). ACTIN (probe: Hs99999903_m1, Life Technologies) was used for normalization purpose. Determinations were performed in triplicate and Relative Quantification (RQ) levels were calculated by ΔΔCt method. n/s = not significant. d Scheme representing ENG coding exons (gray) and translated regions (dark gray). The position of physiological and alternative ATG codons are indicated