FIG 12.

Enhanced Src complex formation with NS/LS-associated Shp2 mutants. (A) HEK-293 cells were transiently transfected either with the Shp2 WT and the indicated Shp2 mutants. Cell lysates were immunoprecipitated (IP) with anti-c-Src antibodies, and immune complexes were immunoblotted (IB) with anti-Shp2 and -Src antibodies. The graph represents the means ± SEM of the densitometric analysis from three independent experiments. Statistical significance was derived using a Dunnett's test comparing Shp2 mutants with the WT. *, P < 0.05; **, P < 0.01. (B) HEK-293 cells were cotransfected with the Shp2 WT or E76A or Y279C mutant and either empty vector (vector), WT human PZR, or the PZR 2YF mutant. Cell lysates were immunoblotted with anti-pPZR(Y241 or Y263) or -Shp2 antibodies. ERK1/2 was used as a loading control. Immune complexes were immunoblotted with anti-Src, -Shp2, and -PZR antibodies.