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Figure 5


Fig. 5. Expression of SHP-2 Y63F and Y279F mutants increases EGF-induced sustained ERK phosphorylation in 293T cells. (A,C) 293T cells were transfected with plasmids encoding wild-type or mutant forms of SHP-2 (10 µg), Gab1 (4 µg) and HA-tagged ERK2 (1 µg), cells were serum-starved, stimulated with EGF for the indicated times, and lysates were probed with phospho-ERK1/2 antibody. Only HA-tagged ERK2, which migrates slower than endogenous ERK1/2, is shown. Blots were stripped and reprobed with HA antibody. (B,D) Values for three unstimulated (as in A) or four EGF-stimulation experiments (as in C). Levels of phosphorylated HA-ERK2 relative to total HA-ERK2 for cells expressing SHP-2 mutants were expressed as a percentage of HA-ERK2 phosphorylation observed in cells expressing wild-type SHP-2. Values are mean ± s.e.m. *P≤0.05, **P≤0.03, ***P≤0.001, t-tests.