Diagram of methodology for mRNA splicing analysis by RT-PCR.

A forward primer in the first exon and alternative reverse primers in the first intron or the fifth exon amplified products of different sizes, as indicated below the figure (bp = base pairs). The spliced transcript was amplified using a forward primer (FP) corresponding to gp91-phox cDNA exon 1 nucleotides 28→52 (5′→3′) and a reverse primer (RP) corresponding to exon 5 nucleotides 407→381 (3′→5′). The unspliced product was detected by the same forward primer, coupled with a reverse primer corresponding to intron 1 nucleotides 226→200 (3′→5′).