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Fig. 4. (A) Cell lines expressing wild-type or 679-AA-mutant receptors were stimulated with EGF for the times indicated. EGFR immunoprecipitates resolved on a 5% acrylamide gel were transferred to nitrocellulose filters and immunoblotted with a ubiquitin-specific antibody. Solid-line brackets indicate location of ubiquitylated EGFR (high molecular mass), arrow indicates 170-kDa EGFR (non-ubiquitylated). (B) The filter in (A) was re-probed with an EGFR-specific antibody. Arrow indicates 170-kDa EGFR (non-ubiquitylated). High molecular mass ubiquitylated EGFR (dashed-line bracket) was not seen unless the film was overexposed (data not shown).





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