Genetic studies of the proband. (A) Schematic illustration of GLRX5 gene. The arrows indicate primers used for genomic amplification (A1-A2, B1-B2, C1-C2), for semiquantitative determination of cGLRX5 (D1-E2), and for exon-intron junction amplification (D1-D2, E1-E2) of proband and control cDNA. (B) Electropherograms of control (C) and proband (P) DNA in the region encompassing the exon 1–intron 1 boundary. The A→G homozygous mutation is boxed. (C) RT-PCR of GLRX5 cDNA from P and controls (C1, C2) PBMC and a negative control (−). From top to bottom: The fragments corresponding to correctly spliced cDNA, obtained by using primers D1-E2, are reduced in the proband. The genomic regions encompassing exon1-intron1 (primers D1-D2) and intron1-exon2 (primers E1-E2) boundaries amplified on cDNA are significantly increased in the proband, suggesting a splicing defect. The identity of the fragments was verified by sequencing. G6PDH was used as control of the reverse transcription reaction and to demonstrate lack of genomic contamination. (D) Relative gene expression of GLRX5 by qRT-PCR showing significant decrease in proband compared to controls. Error bars represent the standard error.