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Figure 5


Fig. 5. Src family kinases are required for netrin to induce UNC5 tyrosine phosphorylation. (A) Netrin fails to induce UNC5 tyrosine phosphorylation in Src/Fyn/Yes triple-knockout MEF cells. UNC5 was transfected with DCC into wild-type and Src/Fyn/Yes triple-knockout MEF cells. 48 hours after transfection, cells were serum-starved for 8 hours and netrin was added for the indicated times. Tyrosine phosphorylation of immunoprecipitated UNC5 was determined. (B) Expression of Src restores netrin-induced UNC5 tyrosine phosphorylation in Src/Fyn/Yes triple-knockout cells. UNC5 protein from transfected cells was immunoprecipitated and subjected to immunoblot with anti-phosphotyrosine antibody. Transfections of Src, UNC5 and DCC are indicated.