Figure 4.
cCbl phosphorylation.
Purified naïve, adaptively tolerant, or preactivated 5C.C7 T cells in (A) or control and anergized A.E7 T cells in (B) were stimulated for 1 min with varying concentrations of anti-TCRβ-biotin mAb (0, 1, or 10μg/ml) and anti-CD4-biotin mAb at 10 μg/ml, followed by cross-linking with streptavidin. Samples were lysed with 1% NP40 buffer, immunoprecipitated with anti-cCbl antiserum, and run on SDS-PAGE. Western blots were performed and probed with anti-phosphotyrosine mAb and then reprobed with anti-cCbl mAb.