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. 2014 Jun 13;289(33):23112–23122. doi: 10.1074/jbc.M114.549550

FIGURE 5.

FIGURE 5.

Nck-1 promotes ELMO1/Dock180-induced Rac1 activity. A, Myc-ELMO1 and Flag-tagged Dock180 were transfected into HEK293T cells with WT Nck-1 or R308K mutant of Nck-1, respectively. 24 h after transfection, the amount of GTP-bound Rac1 was determined as described under “Experimental Procedures.” C, HEK293T cells were transfected with Nck-1 siRNA alone, or together with siRNA-resistant WT Nck-1, or R308K plasmid, respectively. 24 h after transfection, the amount of GTP-bound Rac1 was determined as described in the method. If necessary, empty vector was added to ensure that equal plasmid amounts were transfected in each condition. B and D, graphic representation of the ratio of GTP-Rac1 to total Rac1. Data are expressed as mean ± S.D. of three independent experiments. B, *, p < 0.05 versus untransfected cells. #, p < 0.05 versus ELMO1 and Dock180 co-transfected cells. D, *, p < 0.05 versus untransfected cells. #, p < 0.05 versus Nck siRNA transfected cells.