Skip to main content
. 2014 Feb 28;289(15):10431–10444. doi: 10.1074/jbc.M113.541839

FIGURE 2.

FIGURE 2.

BCAR3 and the related NSP3 induce formation of membrane ruffles, which requires complex formation with BCAR1. a, MCF7 cell populations stably transduced with lentiviral vectors encoding ZsGreen alone (V) or together with wild-type BCAR3 (WT), the BCAR3 R748E single mutant (748), or the BCAR3 L744E/R748E double mutant (744/748) were stained with phalloidin to label filamentous actin (top row) or with an anti-BCAR3 antibody (bottom row). The phalloidin-stained cells were also imaged for ZsGreen fluorescence to identify the cells expressing BCAR3 and ZsGreen from the bicistronic transcript (middle row). BCAR3 wild-type and the R748E single mutant both promote the formation of membrane ruffles compared with the vector control cells (arrows point to examples of ruffles), whereas the BCAR3 L744E/R748E double mutant does not. Furthermore, BCAR3 immunoreactivity is evident in the ruffles. Scale bars = 25 μm. b, the histogram shows the percentage of ZsGreen-expressing cells that contain ruffles. **, p < 0.01, and ***, p < 0.001 for the comparison with vector control-infected cells by one-way ANOVA and Dunnett's post hoc test. c, overexpression of BCAR3 and NSP3 in MCF7 cells promotes the formation of membrane ruffles containing BCAR1. Levels and subcellular localization of endogenous BCAR1 are shown by immunolabeling (arrows point to examples of ruffles). ZsGreen fluorescence identifies the cells expressing BCAR3 or NSP3 from the bicistronic transcripts. Scale bar = 40 μm.