Figure 7

BCAR3 requires p130Cas to antagonize transforming growth factor β signaling. (a) SCP2 cells were transfected with BCAR3 small interfering RNA (siRNA), starved overnight and stimulated with 200 pM transforming growth factor β (TGFβ). Smad2/3 was immunoprecipitated (IP), and levels of p130Cas in the precipitant were examined by Western blot analysis. (b) and (c) SCP2 cells were transfected with BCAR3 siRNA or p130Cas siRNA or together, then starved, stimulated with 200 pM TGFβ and subjected to IncuCyte cell migration assays. Real-time migration profiles were compared for mock-transfected cells and cells transfected with p130Cas siRNA (b) and for cells transfected with p130Cas siRNA and both siRNAs (c). Error bars show standard error of the mean for six biological replicates of a representative experiment (n = 3). (d) The effects of TGFβ on cell migration at the endpoint (36 hours) are plotted as fold changes of relative wound density. Error bars show standard error of the mean. An asterisk indicates statistical significance (*P < 0.05) as determined by unpaired Student’s t-test.