Fig. 2. CytD and LatA transiently activate RhoA, Rac1 and Cdc42. K562-ALCAM cells (A) and KG1 cells (B) in suspension were treated with 2.5 µg/ml CytD for the indicated time. Subsequently, cell lysates were incubated with GST-PAK-CD or GST-C21 and the bound RhoA, Rac1 and Cdc42 molecules were detected by western blot with the respective antibodies. Total lysates served as a control for analyzing total amounts of RhoA, Rac1 or Cdc42 on western blot using the respective antibodies. Similar observations were made using parental K562 cells (data not shown).