Stimulation via anti-TNP TPCR induces coactivation of signal I (ERK1/2) and signal II (PI3K/c-Akt) pathways. (A) Purified T cells were stimulated with immobilized antibodies or TNP for 15 minutes. Cell lysates were resolved by 10% SDS-polyacrylamide gel electrophoresis followed by immunoblotting using phospho-ERK1/2-specific antibody (top). Stripped membranes were then reacted with ERK1/2-specific antibody (bottom). (B) As in panel A, using phospho-c-Akt–specific antibody (top) and c-Akt–specific antibody (bottom) after stripping.