EBP and EEN inhibit the transformation potential of Ras and Ras-mediated activation of Elk-1. (A) Representative colonies of soft agar assays. Ras+EBP+EEN formed small colonies (experiment I), while Ras+EBP+NLSEEN formed large colonies similar to Ras-transformed cells (experiment II). (B) Expression of exogenous EBP-Myc, EEN-FLAG, and NLSEEN-FLAG proteins in stably transfected R6 cells was confirmed by Western blot analysis. (C) EBP and EEN inhibit Ras-mediated activation of Elk-1. RasQ61L-induced activation of Elk-1 (vector control and f; P < .001, ANOVA). Either EBP or EEN alone reduces Elk-1 activation (i and ii; P < .001, ANOVA). The inhibition is further enhanced with both EBP and EEN (ii and iii; P < .001, ANOVA). (D) Dosage-dependent inhibition of EBP and EEN on the Elk-1 transactivation by Ras. Asterisk indicates the concentrations under which significant inhibition on Elk-1 transactivation was observed (P < .05, ANOVA). Error bars represent SDs of triplicate transfections of 2 independent experiments.