Figure 6

Knocking down BCAR3 promotes transforming growth factor β–induced gelatin matrix digestion. (a) SCP2 cells were transfected either with 50 pM scrambled control siRNA (panels 1 and 2), or with 50 pM BCAR3 siRNA (panels 3 and 4), or they were mock-transfected (panels 5 and 6) for 48 hours. Cells were then seeded onto coverslips coated with Alexa Fluor 488–conjugated gelatin, treated with or without 200 pM transforming growth factor β (TGFβ) and allowed to digest the matrix for 36 hours. Representative images show immunofluorescence staining of BCAR3 (white), actin filaments (red) and gelatin matrix (green). The results represent two independent experiments. (b) The total areas of digestion (pixel count) were quantified for ten cells in each condition in seven to ten LSM780 confocal microscopic image files from the two experiments. Error bars show SEM, and an asterisk indicates statistical significance (*P < 0.05) determined by unpaired Student’s t-test. (c) Western blot showing the effect of BCAR3 siRNA using total cell lysates of the same pool of cells used for the gelatin degradation assay.