Figure 8.

Partial rescue of the native proinsulin disulfide bond formation that is perturbed in MIDY. 293T cells transfected with lose-A6/A11 bearing the B22Q MIDY mutation were pulse labeled with ³⁵S-amino acids for 30 min and either untreated (control) or treated with 1 mmol/L diamide for 5 min (diamide) and chased for the times indicated. After washing of cells in ice-cold PBS containing 20 mmol/L NEM, cells were lysed in the presence of 2 mmol/L NEM and immunoprecipitated with anti-insulin. Both cell lysates (C) and media (M) were analyzed by nonreducing (upper) and reducing (lower) Tris–tricine–urea–SDS-PAGE and autoradiography.