Figure 3

Representative Ethidium-bromide stained agarose gel visualising PCR amplicons derived from 17 archival biopsy specimens (lane 1 to 17). A) Quality control of total RNA (free of contaminating genomic DNA), and B) single-stranded cDNA by means of β-actin PCR amplification after DNase treatment. C) Visualised amplicons from nested CCK2R, D) gastrin, and E) CCK2i4svR specific PCR assays as described in methods. The position of the 500 bp size marker in each 100 bp ladder (lane M) is indicated. Black arrows in the left margin indicate the position of a) amplicons derived from genomic DNA or cDNA retaining introns, b) wild-type CCK2 receptor and gastrin mRNA amplicons, and c) CCK2i4svR splice variant amplicons derived from ss-cDNA. White arrows (lane 6, 10, and 17) indicate confirmed CCK2 receptor splice variants by DNA sequencing. Stripped arrows indicate the position of the CCK2i4svR amplicon in sample No. 15. In all amplification assays human genomic DNA (+) was included as control. (-) NTC (no DNA template added to the master mix).