FIG. 2.
Tec and Itk are up-regulated in activated T cells and in Th1 and Th2 effector cells. For T-cell activation, C57BL/6 mouse T cells were isolated as described in the legend to Fig. 1A and stimulated with plate-bound anti-CD3 MAb and soluble anti-CD28 MAb. Cells were harvested at the indicated time points, and lysates were Western blotted for Tec, Itk, and actin. For generation of Th1 and Th2 cells, T cells were sorted for CD4+ CD62hi naïve cells to over 98% purity. T cells were stimulated with plate-bound anti-CD3 MAb and soluble anti-CD28 and differentiated under Th1 or Th2 polarizing conditions for 6 days, and then cells were lysed and Western blotted for Tec, Itk, and actin. Quantitation of Tec and Itk bands is shown after correcting for actin. The data are representative of three experiments.