FIG. 8.

Monoubiquitination of the Met Y1003F receptor leads to its degradation and restores HGF-induced Hrs tyrosine phosphorylation. (A) T47D cells treated with 100 μg/ml cycloheximide were stimulated with 3 nM HGF for the indicated amount of time. Total cell lysates (TCL) (30 μg) were immunoblotted for Met (DL-21) and Erk. (B) T47D cells were stimulated with 1.5 nM HGF for the indicated times and lysed in TGH buffer. Cell lysates were blotted with phospho-Ser217/221 MEK1/2 and total MEK1/2. (C) HEK293 cells were cotransfected with GFP-Hrs and vector or the indicated CSF-Met constructs. GFP-Hrs and Met proteins were immunoprecipitated and blotted with phosphotyrosine antibodies, then stripped and reblotted for GFP and Met, respectively.