Expression of constitutively active Src suppresses SGEF-mediated promotion of cell migration and RhoG activation.

<p>(A) Motility of HEK293T cells was evaluated by Transwell cell migration assays. HEK293T cells transfected with YFP alone or together with the indicated plasmids were plated in the upper chamber of the filters, and cells that had migrated to the underside of the filters were analyzed at 6 h after plating. Results are expressed as means ± SEM (n = 5) of relative cell migration, with SGEF-WT-transfected cell migration set at 1 (*<i>P</i> < 0.05, ***<i>P</i> < 0.001, <i>t</i>-test). (B) HEK293T cells transfected with YFP alone or together with the indicated plasmids were plated in the upper chamber of the filters, and cells that had migrated to the underside of the filters were analyzed at 6 h after plating. Results are expressed as means ± SEM (n = 4) of relative cell migration, with Ephexin4-WT-transfected cell migration set at 1 (ns, not significant, ***<i>P</i> < 0.001, <i>t</i>-test). (C) Cell lysates from HEK293T cells transfected with indicated plasmids were incubated with GST-ELMO-NT, and bound Myc-RhoG (GST-ELMO-NT pull-down) and total cell lysates were analyzed with antibodies against Myc, Flag, and HA. The amount of bound Myc-RhoG was normalized to the total amount of Myc RhoG in cell lysates, and relative RhoG activity was expressed as fold change relative to the basal activity (None). Data are presented as means ± SEM from four independent experiments (*<i>P</i> < 0.05, ***<i>P</i> < 0.001, <i>t</i>-test).