Figure 4. FAK functions upstream of Src kinase to regulate invadopodia formation. (A) MTLn3 cells transiently transfected with control siRNA (Ctlsi) or FAK siRNA (FAKsi-A and FAKsi-B) were cultured on fibronectin-gelatin coverslips in the presence of vehicle control (−PP2) or 2 µM PP2 (+PP2). Cells were stained with anticortactin antibody (green) and rhodamine-phalloidin (red). (B) Quantification of cortactin- and actin-containing invadopodia is expressed as the mean number of invadopodia per cell. (C) MTLn3 cells expressing c-Src or c-Src–527F were transiently transfected with control siRNA or FAK siRNA, cultured on fibronectin-gelatin coverslips, and stained with anticortactin antibody. (D) Quantification of cortactin-containing invadopodia in c-Src– or c-Src–527F-expressing cell lines is expressed as the mean number of invadopodia per cell. (E) MTLn3 cells expressing v-Src were transiently transfected with control siRNA or FAK siRNA and cultured on fibronectin-gelatin coverslips. Cells were stained with anticortactin antibody (green) and rhodamine-phalloidin (red). Data shown are means ± SEM of three independent experiments. *, P < 0.05 by one-way ANOVA compared with control siRNA (−PP2). Bars, 10 µm.
Figure 4.

FAK functions upstream of Src kinase to regulate invadopodia formation. (A) MTLn3 cells transiently transfected with control siRNA (Ctlsi) or FAK siRNA (FAKsi-A and FAKsi-B) were cultured on fibronectin-gelatin coverslips in the presence of vehicle control (−PP2) or 2 µM PP2 (+PP2). Cells were stained with anticortactin antibody (green) and rhodamine-phalloidin (red). (B) Quantification of cortactin- and actin-containing invadopodia is expressed as the mean number of invadopodia per cell. (C) MTLn3 cells expressing c-Src or c-Src–527F were transiently transfected with control siRNA or FAK siRNA, cultured on fibronectin-gelatin coverslips, and stained with anticortactin antibody. (D) Quantification of cortactin-containing invadopodia in c-Src– or c-Src–527F-expressing cell lines is expressed as the mean number of invadopodia per cell. (E) MTLn3 cells expressing v-Src were transiently transfected with control siRNA or FAK siRNA and cultured on fibronectin-gelatin coverslips. Cells were stained with anticortactin antibody (green) and rhodamine-phalloidin (red). Data shown are means ± SEM of three independent experiments. *, P < 0.05 by one-way ANOVA compared with control siRNA (−PP2). Bars, 10 µm.

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