FIG 8.

Migration of FBP1-kd Huh7.5 cells is drastically reduced. (A) Wound-healing assay with control and FBP-kd cells. Cells grown to 100% confluence were starved by restricting FBS in the medium to 0.1% overnight. A wound-like simulation was done by making a scratch across each well. Cells then were supplemented with medium with 10% FBS. Images were taken at the indicated times (left), and the areas covered by the scratches were quantified by ImageJ software (right). AU, arbitrary units. (B) Real-time migration dynamics also indicated a drastic reduction in the migration of FBP1-kd cells. We used the RTCA DP Xcelligence system to measure the real-time migration of control and FBP1-kd Huh7.5 cells (30). Migration is shown as a change in delta cell index versus time. (C) Expression of cell migration markers in control and FBP1-kd cells. Control and FBP1-kd cells were grown to 50% confluence, starved overnight in 0.1% FBS, and then supplemented with 10% FBS. Cell were grown further for 32 h and lysed, and cell lysates were Western blotted for cortactin, p130Cas, and actin (lanes 1 and 2). To detect the level of the phosphorylated form of cortactin and p130Cas, we carried out IP using antibody against cortactin and p130Cas and immunoblotted them for total cortactin or p130Cas, as well as their phosphorylated forms, using antibody against phosphotyrosine (lanes 3 and 4).