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Fig. 6. Expression of SHP-2-Y279F increases PDGF-induced sustained ERK phosphorylation and proliferation in fibroblasts. (A,B) Lysates from serum-starved, PDGF-stimulated NIH3T3 cells, infected with wild-type or mutant SHP-2 retroviruses, were blotted with phospho-ERK1/2 antibody. Changes in ERK1/2 phosphorylation shown are relative to total ERK1/2 levels obtained from reprobed blots. Different exposures are shown for each time point, so that differences between mutants can easily be seen. Results are expressed as a percentage of wild type. (B) Results from three independent experiments (as in A); values are mean ± s.e.m. *P $≤$ 0.05, **P $≤$ 0.02, ***P $≤$ 0.005, using t-tests. (C,D) Tritiated thymidine incorporation was assessed in infected NIH3T3 cells labeled with tritiated thymidine for 24 hours. (C,D) One representative experiment (C; mean ± s.d.), and (D) a composite of three independent experiments in which tritiated thymidine incorporation was compared in cells expressing SHP-2 mutants and in cells expressing wild-type SHP-2. Values are mean ± s.e.m. (some error bars are too small to be visible). Aliquots of infected cells were lysed and probed with SHP-2 antibody. *P $≤$ 0.05, ***P $≤$ 0.001 using a one-way ANOVA followed by a Bonferroni post-hoc test.

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