Real-time PCR detection of deer mouse transcription factors. Total RNA was extracted from 42 hour cultures of T cells with or without KLH activation and used for cDNA synthesis. Real-time PCR was performed using primers described in Table 1 to detect the expression of transcription factors associated with Th1, Th2 or Treg T cells. The mean CT values and standard deviations from triplicate samples obtained from the instrument were subtracted from the number of cycles performed (50) to determine adjusted CT means for unstimulated (-K) and stimulated (KLH) T cell cultures (A). The adjusted CT mean values were then used to calculated the cycle difference (CD) by subtracting the -K value from the KLH value. The relative template abundance (RTA) was then determined by calculating the log2 value of the CD (2CD) (B).