RAC2 Q61R, RAC2 D63V, RAC2 E62K, and RAC2 G12R mutants bind PAK1 RBD. Hemagglutinin-tagged RAC2 (WT) and mutants were expressed for 16 h in HEK293T cells. HA-RAC2 activation level measurements were assessed by GST–PAK–RBD pull-down assay followed by immunoblotting (IB). (A) RAC2 Q61R mutation was compared with WT and the constitutively active mutant RAC2 Q61E. (B) D57N, D63V, and E62K mutations were compared with WT and the constitutively active mutant RAC2 Q61E. (C) RAC2 G12R mutation was compared with WT and the constitutively active mutants RAC2 G12V and RAC2 Q61E. Data are representative for n = 3. Molecular weight (kD) is indicated in the figure. Source data are available for this figure: SourceData F1.