(A) Control or Tks5 siRNA-treated cells were stimulated with 5 nM EGF for 1 min and stained with N-WASP and cortactin antibodies to identify invadopodium precursors (shown by white arrows).
(B) Quantification of invadopodium precursors (means ± SEM) in control and Tks5 siRNA-treated cells. n = 19 (control siRNA) and 22 (Tks5 siRNA) cells.
(C) Stills from Movie S6 showing invadopodia stability. White arrows in control siRNA panel point to a stable invadopodium, whereas white arrows in Tks5 siRNA panels point to examples of unstable invadopodium precursors.
(D) Invadopodia lifetime histograms for control and Tks5 siRNA-transfected cells. Note the complete absence of stable invadopodia (lifetime > 60 min) in Tks5 siRNA cells. n = 49 (control siRNA) and 60 (Tks5 siRNA) invadopodia.
(E and F) Representative invadopodium trajectories in control (E) and Tks5 siRNA-treated (F) cells. Numbers beside each trajectory indicate lifetime (min).
(G) Quantification of invadopodium motility as net distance per lifetime (means ± SEM). n = 39 (control siRNA) and 49 (Tks5 siRNA) invadopodia.
See also Figure S2.