FIG. 6.

Cdc42 activation potential of the DH mutants in cells. (A) The HA-tagged wild-type Dbl or the DH mutants were cotransfected with HA-tagged Cdc42 in Cos-7 cells. At 48 h posttransfection, cell lysates were subjected to GST-PAK1 affinity precipitation. The coprecipitated Cdc42-GTP was detected by anti-HA Western blotting. A sample that was 10% of the amount of whole cell lysates used for GST-PAK1 incubations was also subjected to anti-HA blotting in parallel. WT, wild type. (B) Quantification of the Cdc42-GTP pull-down assays by densitometry measurement. The amount of Cdc42-GTP coprecipitate for the wild-type Dbl cotransfected cells was treated as 100%. The data represent results from four independent experiments.