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Fig. 1. (A) Parental NR6 cells that lack endogenous EGFRs and NR6 cell lines stably expressing wild-type human EGFR or a mutant receptor with an inactivating 679-AA substitution, were incubated with ¹²⁵I-EGF for 2 hours at 4°C, followed by incubation with a chemical cross-linker at room temperature for 15 minutes. (B) NR6 cell lines expressing wild-type or 679-AA-mutant receptors were pulse-labeled with a mixture of [³⁵S]cysteine and [³⁵S]methionine for 30 minutes followed by a 3-hour incubation in chase medium. The pulse-labeled cells were then stimulated with EGF for 0, 1 or 2 hours and cell lysates were immunoprecipitated with a human EGFR-specific antibody. Equal aliquots of total cellular protein (A) or EGFR immunoprecipitates (B) were resolved by SDS-PAGE for autoradiographic detection. Molecular mass standards: 200,000 Da (myosin); 116,300 Da (ß-galactosidase).

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