FIGURE 6.

Knockdown of mda-9/syntenin suppresses the plasma membrane translocation of ILK, PINCH1, and α-parvin complex during adhesion to COL-I. A, HEK 293T17 cells were transfected with the indicated constructs for 48 h. Whole cell lysates were immunoprecipitated with HA antibody, and then immunoprecipitates (IP) and whole cell lysates (Input) were blotted with the indicated antibodies. B, MCF-7 cells transfected with a control vector or FLAG-tagged mda-9/syntenin vector were plated onto COL-I-coated dishes for the indicated periods of time and then fractionated into cytosol and plasma membrane fraction. Both fractions were blotted with the indicated antibodies. ErbB2 was used as the membrane protein marker, and GAPDH was used as the cytosol protein marker. C, MDA-MB-231 cells were transfected with a control siRNA, or mda-9/syntenin-targeted siRNA were plated onto COL-I-coated or uncoated coverslips for 30 min, fixed, and stained with appropriate primary and secondary antibodies to visualized the endogenous ILK (red, Alexa 546) and PINCH1 (green, Alexa 488). D, MDA-MB-231 cells were prepared as described in c and stained with the appropriate primary and secondary antibodies to visualized the endogenous ILK (green, Alexa 488) and α-parvin (red, Alexa 546). Bar, 20 μm.