The tyrosine dephosphorylation of cortactin is required for stable long-lived invadopodia. (A) Quantification of the average lifetimes of invadopodium precursors in cortactin WT, 3YF, and 3YE GFP-actin cell lines. n = number of invadopodium precursors: WT (133), 3YF (183), 3YE (152) from more than three independent experiments. (B) The fluorescence intensity of Arp2, cofilin, and N-WASp at invadopodium precursors in WT, 3YF, and 3YE cell lines. P values for A and B are compared with WT. n = number of invadopodium precursors: >30, three independent experiments. (C) Pearson's correlation analysis between cortactin and cofilin (r = 0.80) vs. pY421-cortactin/total cortactin and cofilin (r = −0.51) in invadopodium precursors. Error bars represent ± 95% confidence interval. (D) Quantification of the fluorescence intensity of pY421-cortactin/total cortactin at invadopodium precursors in ctrl vs. v-Src expressing cells. (E) Quantification of average invadopodium precursors lifetimes in GFP-actin cells expressing v-Src. (F) Quantification of the average FN degradation area/field, (G) number of invadopodium precursors/cell, and (H) FN degradation area normalized to the number of invadopodium precursors/cell in ctrl vs. v-Src expressing cells.