Fig. 3

A Reverse transcription-PCR analysis of mRNAs from peripheral blood leukocytes of three normal controls (NC1-3) and the patient. In addition to a minor expression of the full length GLA transcript (a: 392 bp), the patient showed two major aberrant GLA transcripts (b: 414 bp and c: 277 bp). The relative ratio of the three transcripts was quantified by means of examing their grey scale using ImageJ (http://imagej.nih.gov/ij/). B qRT-PCR analysis using 2 pairs of cross-exon primers were performed on total RNA from peripheral blood leukocyte of three normal controls and the patient for the quantification of GLA full-length mRNA transcript. Levels were normalized to the amount of GAPDH. Data represented the mean ± SD of triplicate experiments. C Schematic of normal and aberrant splicing patterns. D Sanger sequencing of the full-length 392-bp transcript (a), the 414-bp aberrant transcript (b) and the 277-bp aberrant transcript (c). E: 57-bp pseudoexon; M: molecular weight markers; q: donor-site shift followed by the number of bp that are skipped or included; wt: wild type; △x: skipping of all or part of exon x; ▼: inclusion of intronic sequence in transcript