(A) Schematic of DEPDC1B polypeptides that were used in (B), (C), (D), and (E). The number above the diagram indicates the amino acid residues. (B) Immunoblots showing that GFP-tagged DEPDC1B full-length (lane 1) and fragments (lanes 2 and 3) are expressed in transfected HeLa cells. (C-E) Plasmids encoding MYC-RAC1 (C), MYC-RHOA (D), or MYC-CDC42 (E) were co-transfected into HeLa cells with empty vector or plasmids encoding GFP-DEPDC1B-FL, GFP-DEPDC1B-1-151, or GFP-DEPDC1B-151-529. The transfected cells were subjected to PBD pull-down assays to assess the levels of activated MYC-RAC1 (C) and MYC-CDC42 (E) or subjected to RBD pull-down assays to assess the levels of activated MYC-RHOA (D). Typical immunoblots for the activation assays of RAC1 (Ca), RHOA (Da), and CDC42 (Ea) were shown. Relative amount of activated MYC-RAC1 (Cb), MYC-RHOA (Db), and MYC-CDC42 (Eb) in each group of transfected cells were quantitated based on three independent experiments. The error bars indicate standard deviation around the mean. In (Cb), (Db), and (Eb), FL, 1-151, and 151-529 represent GFP-DEPDC1B-FL, GFP-DEPDC1B-1-151, and GFP-DEPDC1B-151-529, respectively. **, p<0.01; *, p<0.05.