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. 1997 Apr 21;137(2):387–398. doi: 10.1083/jcb.137.2.387

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Stimulation of JNK activity by Tiam1. (A) COS-7 cells were transfected with pCMV-Flag-JNK1 and empty vector (pcDNA-neo), full-length Tiam1 (FL1591), or constitutively activated (V12)Rac1. Cotransfection with dominant-negative (N17)Rac1 was performed as indicated. The activity of immunoprecipitated Flag-JNK was determined 48 h after transfection by an in vitro kinase assay using bacterially expressed GST-Jun as a substrate. The expression of Tiam1, Myc-tagged (N17)Rac, and Flag-tagged JNK was analyzed by Western blotting and is shown underneath the graph. (B) NIH3T3 cells were transfected with pCMV-Flag-JNK1 and mutant Tiam1 constructs. JNK activity was determined as described. All data are presented as fold stimulation of empty vector controls. Equal amounts of expressed Flag-JNK1 and Tiam1 proteins, as determined by Western blotting, were used. Data represent mean ± standard deviation of at least two independent experiments.