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. 2002 Dec;22(24):8648–8658. doi: 10.1128/MCB.22.24.8648-8658.2002

FIG. 6.

FIG. 6.

TCR-induced tyrosine phosphorylation, activation of ERK, JNK, and p38MAPKs, and intracellular calcium fluxes in thymocytes. (A and B) Double-positive thymocytes were incubated with anti-CD3 Ab for the indicated times. Induction of tyrosine phosphorylations of total proteins (A), ERK (B) and JNK (B) in whole-cell lysates was analyzed by immunoblotting. p38MAPK activity was measured using an immunocomplex kinase assay (B). (C) CD4 single-positive thymocytes were incubated with anti-CD3 Ab for the indicated times. Induction of ERK, JNK, and p38MAPK phosphorylation was analyzed by immunoblotting. (D) Intracellular calcium fluxes were measured in double-positive thymocytes or CD4 single-positive thymocytes using Fluo-3. Biotinylated anti-CD3 Ab was first added at 0 s, and then streptavidin was added at 20 s (arrow indicated) for cross-linking.