Figure 5

Decreased Rac1 and RhoA activity in DGK ζ knockdown cells. Assay of global Rac1 and RhoA activity. Detergent extracts from the indicated cell lines were incubated with immobilized GST-PBD or GST-RBD and the bound proteins were analyzed by immunoblotting (IB) for Rac1 (A) or RhoA and tubulin (C). Total Rac1 and RhoA levels in the cell lysates are shown. (B and D) Quantification of active Rac1 and RhoA levels, respectively, by densitometric analysis of immunoblots. The data were normalized to the amount of total Rac1 or RhoA protein and are expressed as a percentage of the activity in SW620 cells. Values are the average of at least three independent experiments ± S.E.M. The asterisks indicate a highly significant difference from SW620 cells (P < 0.005) by Student’s t-test.