Figure. 1.
TRAP recruitment to the human IR in intact cells. (A) CHO-IR cells were serum starved before stimulation with 100 nM insulin for 5 min at 37°C. After a cross-linking reaction with 100 μM BMH, cell lysates were prepared and then immunoblotted with anti-IR β-subunit or anti-IR α-subunit antibodies. (B) CHO-IR cells were labeled with [35S]Met/Cys for 16 h before stimulation with 100 nM insulin for 5 min at 37°C. After a cross-linking reaction, endogenous IR was immunoprecipitated with anti-IR antibodies, resolved by SDS-PAGE, and detected by autoradiography. Right margin, α- and β-subunits of the IR; left margin, Mr × 10−3; asterisk, TRAP/IR β-subunit complex.