Fig. 3. Normal surface activation marker expression and proliferation response of ConA-stimulated 311R and 288R CD4+ T cells. Splenic CD4+ T cells prepared from 6- to 8-week-old mice were stimulated with ConA (5 µg/ml) and cultured under Th1- and Th2-polarizing conditions (see methods), respectively. (A) Cells were stained for CD25 directly after isolation (0 h) and 48 h after ConA stimulation. The experiment was repeated three times with similar results. (B) To determine the proliferation response after ConA stimulation cells were measured in a 24 h-rhythm by the ViaLight® assay. RLUs were measured at indicated time points post-stimulation and normalized to RLUs obtained prior stimulation. The fold amplification rate is plotted. Results shown are representative of three independent experiments.