Figure 2

Real-time PCR analysis of t-PA (a) and plasminogen activator inhibitor 1 (PAI-1) (b) gene expression in Pa and Ao isolated from the same individual, in comparison with glyceraldehyde-3-phosphate dehydrogenase gene expression. Defined dilutions of cDNA were amplified to determine the linear relationship between the threshold cycle (Ct) and the log starting copy number. In all experiments the correlation coefficients were between 0.98 and 0.99. Quantitation relies on the detection of Ct, which represents the cycle when a fluorescence signal exceeds the mean of background fluorescence plus 10 times the standard deviation in cycles 3 to 15. Plotting the log of the starting copy number of template molecules for several standard concentrations versus Ct reveals a linear correlation. The bars show the ratios between t-PA and PAI-1 Cts and glyceraldehyde-3-phosphate dehydrogenase Ct. Arbitrary units of the relative amount of the gene of interest are displayed on the y-axis. The differences illustrated in b were statistically significant. Error bars represent standard deviations.