Analysis of the altered electrophoretic mobility of FVIIArg¹⁰⁰. After a 15-minute pulse with [³⁵S] methionine, stably transfected cells expressing FVIIwt and FVIIArg¹⁰⁰ were chased for 60 minutes and 120 minutes, respectively, to analyze FVII in cell lysates. In the conditioned media, FVIIwt and FVIIArg¹⁰⁰ were both investigated after 180 minutes of chase. FVIIwt (wt) and FVIIArg¹⁰⁰ (100) were immunoprecipitated using a MoAb against FVII. (A) Equivalent amounts of cell lysate were analyzed by 8% SDS-PAGE before (-) or after treatment with Endo H and N-Glycanase (N-Gly). (B) Conditioned media were analyzed before (-) or after treatment with neuraminidase (Neura) by 10% SDS-PAGE. (C) Conditioned media were evaluated under nonreducing conditions (control) or after reduction by 100 mmol/L Dithiothreitol (DTT). The 77 kD band was nonspecific because it was also observed in conditioned media from untransfected CHO cells (data not shown). Molecular weight markers in kD are indicated on the left.