Reduced upregulation of NFAT1 protein in UCB T cells during primary stimulation. (A) MNC from 2 UCB units and 2 adults were stimulated in parallel with Con A for 40 hours. At 0 hour (unstim), 18 hours, and 40 hours of stimulation, T cells were purified and extracted and NFAT1 protein expression analyzed by Western blotting. The results shown are representative of 5 independent experiments, comparing in each experiment 2 UCB and 2 adult controls analyzed together and run on the same gel. (B) Graphic representation of the mean (±SE) of the quantification of relative NFAT1 expression after stimulation with Con A. Gel loading was normalized for each lane with the actin band and the intensity of each NFAT1 band was calculated as relative percentage of the most intense NFAT1 band on each gel. The most intensive band was set arbitrarily at 100% and relative percentages were then averaged and graphed.