RT-PCR analysis of platelet RNA for human and mouse αIIb.

(A) Agarose gel of size-fractionated nonquantitative RT-PCR products for mαIIb (top) or hαIIb (bottom) from total platelet RNA for the various transgenic lines. The mαIIb samples were amplified for 20 PCR cycles within the linear range of amplification for comparison of relative cDNA levels. The hαIIb samples were amplified for 33 PCR cycles, beyond the linear range of amplification, to facilitate detection of hαIIb within all the diversely expressed transgenic lines. Size markers (M) are indicated to the left and the observed cDNA or genomic bands indicated at the right of each figure. The observed genomic bands were variably present and only seen with the 2 lanes indicated where there appears to be little hαIIb message, but high transgene copy number, was present. Copy numbers are listed as numbers above each lane. WT indicates wild-type mouse platelet RNA control; W, water; mαIIb, cDNA plasmid RT amplified; HP,human platelet RNA control. (B) Analysis of semiquantitative RT-PCR comparing relative expression of human to mouse αIIb message versus the copy number for a particular line. The open diamonds are the2.5hαIIb lines, the closed circles are the7.1hαIIb lines, and the closed squares are the3′+hαIIb lines. The experiments were done in triplicate and repeated at least 3 times. A single SD is indicated for each point.