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Figure 1


Fig. 1. WAVE2 undergoes a mobility shift assessed by SDS-PAGE. (A) NIH 3T3 fibroblasts were serum starved and stimulated with 10% serum for the indicated times. Cells were incubated with 10 µM Uo126 or DMSO control where indicated. Lysates were immunoblotted for the indicated protein species. ErkP, phospho-Erk; Erk t, total Erk. (B) Domain structure of WAVE2 and truncation constructs. (C) GST-WAVE2 constructs were purified from Cos-7 cells growing in 10% serum and treated with alkaline phosphatase (AP) where indicated before resolution by SDS-PAGE and silver staining. (D) GST-{Delta}SHD/{Delta}VCA constructs containing the indicated alanine (A) mutations were purified and treated as above. 308A is included as an example of a Ser-to-Ala mutation that does not affect the mobility of WAVE2. Note the disappearance of faint upper species in the 343A mutant. An aliquot of each phosphatase-treated sample was incubated with recombinant Erk2 (E). AAA, 343A–346A–351A triple mutant.





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