Figure 2
Relationship between the IL-12 p40 activity and the amount of the transfected PGK-p40. a) BHK-21 cells plated in 24-well plates at the density of 105/well were transfected with the indicated amount of PGK-p40 plasmid. b) initial copy number of cDNA transcribed (+RT) from cell samples transfected with various amounts of the p40-encoding plasmid. The initial copy numbers were determined by QPCR as described in the methods section by using a FAM-labeled primer set specific for IL-12b (Certified Lux™ primer set 310H-01, Invitrogen). Samples incubated in the absence of RT (-RT) produced no significant amplification and were omitted from the calculations. The Ct (dRn) values of the samples were plotted against a standard curve of known copy numbers of p40-encoding plasmid run in parallel (RSq = 0.982) to obtain the initial copy number values.